About us. Contacts & Ordering. highqu assists life science by fueling it with research tools of premium quality.

Transcription

1

2 About us highqu assists life science by fueling it with research tools of premium quality. Our dedication to the molecular biology field has enabled us to deliver outstanding reagents to the life science market. In our commitment to science, we combine excellence with simplicity in order to provide the solutions that scientists constantly seek. We support researchers whose professionalism makes science come alive. highqu honors people who have a higher respect for humanity itself than for anything invented by humans. We are devoted to deliver intelligent products and services that render a better life for all. This is our interpretation of life science: we are people, we serve life. Professionally simple We offer premium tools for researchers We keep our word We contribute to your success Your highqu team Contacts & Ordering HEAD OFFICE highqu GmbH Nelkenstrasse Kraichtal Germany T: F: info@highqu.com ORDERING T: F: order@highqu.com TECHNICAL SUPPORT T: F: tech-support@highqu.com SALES T: F: sales@highqu.com

3 Content Product Index...2 qpcr Selection: Instrument Compatibility of Probe and Dye High Resolution Melting Analysis Selection: NEW...9 One Step RT qpcr Selection: Instrument Compatibility for Probe and Dye-based 1Step RT qpcr Kits Step RT qpcr Probe Kit Step RT qpcr Probe ROX L Kit Step RT qpcr Probe ROX H Kit Step RT qpcr Green ROX L Kit Step RT qpcr Green ROX H Kit End-point PCR Selection: Standard, Hot-start, Long, High fidelity, Direct PCR NEW Taq DNA Polymerase RPH Mastermix, 2X RT PCR & Reverse Transcription Selection Step RT PCR Kit Reverse Transcriptase - NEW NEW NEW PCR Reagents dntp Sets & Mixes PCR Water Cloning - NEW NEW Electrophoresis DNA Electrophoresis Ladders Protein Electrophoresis Ladders order@highqu.com Order Tel:

4 Product Index CAT.# PRODUCT CONCENTRATION SIZE SIZE UNIT PAGE QPP0101 2X 200 r of 20 µl 4 QPP0105 qpcr Probe Mix 2X 1000 r of 20 µl QPP0201 Rox L Mix 2X 200 r of 20 µl 5 QPP0205 2X 1000 r of 20 µl QPP0301 qpcr Probe Rox H Mix 2X 200 r of 20 µl 6 QPP0305 qpcr Probe Rox H Mix 2X 1000 r of 20 µl QPD0101 2X 200 r of 20 µl 7 QPD0105 2X 1000 r of 20 µl QPD0201 2X 200 r of 20 µl 8 QPD0205 2X 1000 r of 20 µl QPD0301 NEW 2X 200 r of 20 µl 9 QPD0305 NEW Mix 2X 1000 r of 20 µl QOP0101 1Step RT qpcr Probe Kit 2X & 20X 200 r of 20 µl 11 QOP0105 1Step RT qpcr Probe Kit 2X & 20X 1000 r of 20 µl QOP0201 1Step RT qpcr Probe Rox L Kit 2X & 20X 200 r of 20 µl 12 QOP0205 1Step RT qpcr Probe Rox L Kit 2X & 20X 1000 r of 20 µl QOP0301 1Step RT qpcr Probe Rox H Kit 2X & 20X 200 r of 20 µl 13 QOP0305 1Step RT qpcr Probe Rox H Kit 2X & 20X 1000 r of 20 µl QOD0101 1Step RT qpcr Green Rox L Kit 2X & 20X 200 r of 20 µl 14 QOD0105 1Step RT qpcr Green Rox L Kit 2X & 20X 1000 r of 20 µl QOD0201 1Step RT qpcr Green Rox H Kit 2X & 20X 200 r of 20 µl 15 QOD0205 1Step RT qpcr Green Rox H Kit 2X & 20X 1000 r of 20 µl PCE u/µl 500 u 17 PCE u/µl 2500 u PCM0201 NEW 2X 200 r of 50 µl 18 PCM0205 NEW 2X 1000 r of 50 µl PCM0101 2X 200 r of 50 µl 19 PCM0105 2X 1000 r of 50 µl PCE0201 Taq DNA Polymerase 5 u/µl 1500 u 20 PCE0202 Taq DNA Polymerase 5 u/µl 3000 u HSE u/µl 500 u 21 HSE u/µl 2500 u HSM0201 2X 200 r of 50 µl 22 HSM0205 2X 1000 r of 50 µl HLE u/µl 250 u 23 HLE u/µl 1250 u HLM0101 2X 200 r of 50 µl 24 HLM0105 2X 1000 r of 50 µl HLE u/µl 200 u 25 HLE u/µl 1000 u RTK0201 1Step RT PCR Kit 2X & 20X 100 r of 50 µl 27 RTE0301 NEW Reverse Transcriptase 200 u/µl u 28 RTE0305 NEW Reverse Transcriptase 200 u/µl u RTK0101 NEW q 5X & 20X 25 r of 20 µl 29 RTK0104 NEW q 5X & 20X 100 r of 20 µl RNI0201 NEW RNase Inhibitor 40 u/µl 3000 u 30 RNI0202 NEW RNase Inhibitor 40 u/µl u NUM mm dntp Mix 25 mm each 1 ml 31 NUM mm dntp Mix 10 mm each 1 ml 31 NUS mm dntp Set 100 mm 4 x 0.25 ml 31 WAT0110 PCR Water 10 x 1 ml 32 RLK0101 NEW Rapid Ligation Kit 1 r/µl 40 r of 20 µl 33 RLK0105 NEW 1 r/µl 200 r of 20 µl HER0101 NEW 1 r/µl 40 r of 25 µl 34 HER0105 NEW 1 r/µl 200 r of 25 µl DNL0102 Ladder 5 µl/appl. 200 minigel appl. (5 µl) 35 DNL bp DNA Ladder 5 µl/appl. 200 minigel appl. (5 µl) 35 DNL bp DNA Ladder 5 µl/appl. 200 minigel appl. (5 µl) 35 DNL µl/appl. 200 minigel appl. (5 µl) 35 PRL µl/appl. 200 minigel appl. (5 µl) 36 PRL µl/appl. 200 minigel appl. (5 µl) 36 2 Info Tel: info@highqu.com

5 qpcr Selection: Instrument Compatibility of Probe and Dye-based qpcr Mastermixes Probe-based qpcr Green dye-based qpcr Probe Mix, Probe ROX L Probe ROX Instruments Green ROX L Green ROX H 2X Mix, 2X H Mix, 2X Mix, 2X Mix, 2X page 4 page 5 page 6 page 7 page 8 Applied Biosystems: 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOnePlus Applied Biosystems: 7500, 7500 FAST, Viia7, QuantStudio 12K Flex Agilent (Stratagene):, Mx3000P, Mx3005P, Mx4000P BioRad: icycler, MyiQ, iq5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Analytic Jena: qtower Cepheid: SmartCycler Eppendorf: Mastercycler ep realplex, Mastercycler realplex 2S Illumina: Eco QIAGEN (Corbet): Rotor-Gene Q, Rotor- Gene 3000, Rotor-Gene 6000 Roche Applied Science: LightCycler 480, LightCycler Nano Takara: Thermal Cycler Dice Techne: Quantica High Resolution Melting Analysis Selection: Instrument Compatibility of qpcr HRM Mix (page 9) Applied Biosystems: BioRad: Eppendorf: Illumina: Roche Applied Science:, QuantStudio 12K Flex Mastercycler ep realplex Mastercycler realplex 2S Eco QIAGEN: Rotor-Gene Q, Rotor-Gene 6000 LightCycler 480, LightCycler Nano order@highqu.com Order Tel:

6 QPP r of 20 µl 2 x 1 ml - 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized buffer, QPP r of 20 µl 10 x 1 ml - ROX is not included. 10 x 1 ml - PCR Water Storage: In the dark at -20 C. qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of gdna, cdna, viral DNA, low copy number genes, gene expression analysis Universal - both standard and fast cycling, all probe qpcr assays, GC or AT rich templates Excellent for both single-plex & multiplexing Rapid extension, early Ct Supplied with PCR Water for maximum convenience highqu qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates, in multiplexing and guaranty rapid extension with early Ct values with minimum or no optimization. Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 3. PERFORMANCE copies of the target: TaqMan probe amplification traces from plasmid dilution series of 1x10 6 copies to 10 copies of DNA. 95 o C 2 m, 40 x 95 o C 10 s & 60 o C 15 s, Biorad CFX. Human gene ACVR2B. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe cdna Template or gdna Template PCR Water qpcr Mix, 2X 200 nm final c. (0.4 µl of 10 µm) <100 ng or 1 g to 10 l 10 l Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation Denaturation Anneal./extension 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna 40 cycles: 95 C - 5 sec 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. 4 Info Tel: info@highqu.com

7 ROX L Mix, 2X QPP r of 20 µl 2 x 1 ml - ROX L Mix, 2X 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized buffer, QPP r of 20 µl 10 x 1 ml - ROX L Mix, 2X 10 x 1 ml - PCR Water low ROX concentration. Storage In the dark at -20 C. qpcr on instruments calibrated with low ROX conc. qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of gdna, cdna, viral DNA, low copy number genes, gene expression analysis Universal - both standard and fast cycling, all probe qpcr assays, GC or AT rich templates Excellent for both single-plex & multiplexing Rapid extension, early Ct Supplied with PCR Water for maximum convenience highqu qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates, in PERFORMANCE copies of the target: TaqMan probe amplification traces from plasmid dilution series of 1x10 6 copies to 10 copies of DNA. 95 o C 2 m, 40 x 95 o C 10 s & 60 o C 15 s, Biorad CFX. Human gene LIMK1. multiplexing and guaranty rapid extension with early Ct values with minimum or no optimization. Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 3. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe cdna Template or gdna Template PCR Water qpcr Mix, 2X 200 nm final c. (0.4 µl of 10 µm) <100 ng or 1 g to 10 l 10 l Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation Denaturation Annealing/extension 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna 40 cycles: 95 C - 5 sec 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. order@highqu.com Order Tel:

8 ROX H Mix, 2X 2 x 1 ml - ROX H Mix, 2X QPP r of 20 µl 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized 10 x 1 ml - ROX H Mix, 2X buffer, high ROX concentration. QPP r of 20 µl 10 x 1 ml - PCR Water Storage In the dark at -20 C. qpcr on instruments calibrated with high ROX conc. qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of gdna, cdna, viral DNA, low copy number genes, gene expression analysis Universal - both standard and fast cycling, all probe qpcr assays, GC or AT rich templates Excellent for both single-plex & multiplexing Rapid extension, early Ct Supplied with PCR Water for maximum convenience highqu qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates, in multiplexing and guaranty rapid extension with early Ct values with minimum or no optimization. Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 3. PERFORMANCE copies of the target: TaqMan probe amplification traces from plasmid dilution series of 1x10 6 copies to 10 copies of DNA. 95 o C 2 m, 40 x 95 o C 10 s & 60 o C 15 s, Biorad CFX. Human gene ACVR1B. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe cdna Template or gdna Template PCR Water qpcr Mix, 2X 200 nm final c. (0.4 µl of 10 µm) <100 ng or 1 g to 10 l 10 l Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation Denaturation Anneal./extension 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna 40 cycles: 95 C - 5 sec 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. 6 Info Tel: info@highqu.com

9 Green ROX L Mix, 2X QPD r of 20 µl QPD r of 20 µl Storage In the dark at -20 C. 2 x 1 ml - Green ROX L Mix, 2X 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized buffer, 10 x 1 ml - Green ROX L Mix, 2X low ROX concentration. 10 x 1 ml - PCR Water qpcr from gdna, cdna, viral DNA, low copy number genes Relative gene expression analysis, absolute quantification qpcr on instruments calibrated with low ROX conc. qpcr assays based on fluorescence of intercalating dye Universal - standard and fast cycling, GC or AT rich templates Highest sensitivity, rapid extension, early Ct values Supplied with PCR Water for maximum convenience highqu qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the Green Mixes are available in different versions with low or high ROX concentration. See the selection table on page 3. PERFORMANCE Green Mix (blue curves) provides in many cases earlier Ct values compared to competitor mastermixes. Conditions: 95 o C 2 m, 40 x 95 o C 10 s & 60 o C 15 s, Roche LightCycler 480. Amplification of mouse ACTG1 from cdna dilution series. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. cdna Template or <100 ng or gdna Template 1 g PCR Water to 10 l qpcr Mix, 2X 10 l Mix gently, avoid bubbles. Place into the instrument (SYBR Green or FAM channel), set like: Initial denaturation 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. order@highqu.com Order Tel:

10 Green ROX H Mix, 2X QPD r of 20 µl QPD r of 20 µl Storage In the dark at -20 C. 2 x 1 ml - Green ROX H Mix, 2X 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized buffer, 10 x 1 ml - Green ROX H Mix, 2X high ROX concentration. 10 x 1 ml - PCR Water qpcr on instruments calibrated with high ROX conc. qpcr assays based on fluorescence of intercalating dye Quantification of gdna, cdna, viral DNA, low copy number genes, gene expression analysis Universal - both standard and fast cycling, GC or AT rich templates Highest sensitivity, rapid extension, early Ct Supplied with PCR Water for maximum convenience highqu qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the qpcr Green Mixes are available in different versions with low or high ROX concentration. See the selection table on page 3. PERFORMANCE compared to competitor mastermixes. Conditions: 95 C 2 m, 40 x 95 C 10 s & 60 C 15 s, Roche LightCycler 480. Amplification of mouse ACTG1 from cdna dilution series. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. cdna Template or <100 ng or gdna Template 1 g PCR Water to 10 l qpcr Mix, 2X 10 l Mix gently, avoid bubbles. Place into the instrument (SYBR Green or FAM channel), set like: Initial denaturation 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. 8 Info Tel: info@highqu.com

11 HRM Mix, 2X - NEW QPD r of 20 µl QPD r of 20 µl Storage In the dark at -20 C. 2 x 1 ml - HRM Mix, 2X 2 x 1 ml - PCR Water Hot Start qpcr components: dntps at 0.25 mm, optimized buffer, 10 x 1 ml - HRM Mix, 2X proprietary saturating intercalating dye 10 x 1 ml - PCR Water High Resolution Melting analysis (HRM): Detection of sequence variations SNP genotyping Methylation analysis Mutation scanning Time and costs saving analysis of sequence variations Universal - standard or fast cycling, GC or AT rich templates Highest sensitivity, no optimization required Supplied with PCR Water for maximum convenience High Resolution Melting analysis (HRM) is a fast and simple technique for identification of DNA sequence variations. It allows identifying single nucleotide differences by detecting minor changes in qpcr melting curves. highqu HRM qpcr Mix includes a proprietary intercalating saturating dye showing no inhibition for PCR. The dye has the same affinity for both AT or GC rich sequences what leads to highest accuracy in genotyping. The hot-start function in the mix is based on the small molecular inhibitor technology and allows achieving highest sensitivity and specificity under both standard and fast qpcr cycling conditions. The mix provides excellent performance on both AT and GC rich templates and reliable results with minimum or no optimization. COMPATIBILE INSTRUMENTS Applied Biosystems BioRad Eppendorf Illumina 7500, 7500 FAST, 7900, 7900HT FAST, 7, QuantStudio 12K Flex Mastercycler ep realplex Mastercycler realplex 2S Eco QIAGEN Rotor-Gene Q, Rotor-Gene 6000 Roche Applied Science LightCycler 480, LightCycler Nano Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. cdna Template or <100ng or gdna Template <1 g PCR Water to 10 l HRM Mix, 2X 10 l Mix gently, avoid bubbles. Place into the instrument (SYBR Green or FAM channel), set like: Initial denaturation 1 cycle: 95 C - 2 min for cdna, or 1 cycle: 95 C - 3 min for gdna Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. order@highqu.com Order Tel:

12 One Step RT qpcr Selection: Instrument Compatibility for Probe and Dye-based 1Step RT qpcr Kits Probe-based One-step RT qpcr Green dye-based One-step RT qpcr 1Step RT qpcr Probe 1Step RT qpcr Probe 1Step RT qpcr Probe Instrument 1Step RT qpcr Green 1Step RT qpcr Green Kit ROX L Kit ROX H Kit ROX L Kit ROX H Kit page 11 page 12 page 13 page 14 page 15 Applied Biosystems: 7000, 7300, 7700, 7900, 7900HT, 7900HT FAST, StepOne, StepOnePlus Applied Biosystems: 7500, 7500 FAST, Viia7, QuantStudio 12K Flex Agilent (Stratagene):, Mx3000P, Mx3005P, Mx4000P BioRad: icycler, MyiQ, iq5, Opticon, Opticon 2, Chromo4, MiniOpticon, CFX96, CFX384 Analytic Jena: qtower Cepheid: SmartCycler Eppendorf: Mastercycler ep realplex, Mastercycler realplex 2S Illumina: Eco QIAGEN (Corbet): Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000 Roche Applied Science: LightCycler 480, LightCycler Nano Takara: Thermal Cycler Dice Techne: Quantica 10 Info Tel: info@highqu.com

13 1Step RT qpcr Probe Kit 2 x 1 ml - 1Step RT qpcr Probe Mix, 2X QOP r of 20 µl 1 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 2 x 1 ml - PCR Water 10 x 1 ml - 1Step RT qpcr Probe Mix, 2X QOP r of 20 µl 5 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 10 x 1 ml - PCR Water Storage In the dark at -20 C. Hot Start Taq, dntps at 0.25 mm, optimized buffer 20X concentrated blend of modified MMuLV RT and RNase Inhibitor. RT qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of any RNA template (mrna, total RNA, viral RNA), low copy number genes highqu 1Step RT qpcr mastermixes in combination with a blend of thermostable and extremely active Reverse Transcriptase & advanced RNase Inhibitor (RT Mix) allow for a single step one tube RT qpcr. qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. To suit the broad instrument range the 1Step RT qpcr Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 10. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control, no RT Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Higher amounts of RT3 Mix improve Ct, but primer dimers may appear. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Reverse transcription & qpcr in one tube with highest sensitivity Efficient cdna synthesis ensured by the thermostable Reverse Transcriptase & advanced RNase Inhibitor blend Universal Kit for standard and fast cycling, GC/AT rich templates Rapid extension, early Ct Supplied with PCR Water for maximum convenience PRECAUTIONS FOR WORK WITH RNA Take care to prevent RNA from degradation by widely spread and stable RNases. Prepare crude samples and set up reactions in different dedicated areas, use DEPC-treated nuclease-free labware and gloves. Before the cdna synthesis, check RNA quality on denaturing agarose gel to be sure you have good quality material. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe 200 nm final c. (0.4 µl of 10 µm) Total RNA template or 1 pg to 1 µg or mrna template >0.01 pg PCR Water to 10 l 1Step RT qpcr Mix, 2X 10 l RT3 Mix, 20X 1-2 l Mix gently, avoid bubbles. Place into the instrument set like: Reverse Transcription 1 cycle: C - 10 min Initial denaturation 1 cycle: 95 C - 2 min Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. order@highqu.com Order Tel:

14 1Step RT qpcr Probe ROX L Kit 2 x 1 ml - 1Step RT qpcr Probe ROX L Mix, 2X QOP r of 20 µl 1 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 2 x 1 ml - PCR Water 10 x 1 ml - 1Step RT qpcr Probe ROX L Mix, 2X QOP r of 20 µl 5 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 10 x 1 ml - PCR Water Storage In the dark at -20 C. Hot Start Taq, dntps at 0.25 mm, optimized buffer, low ROX concentration. 20X concentrated blend of modified MMuLV RT and RNase Inhibitor. qpcr on instruments calibrated with low ROX conc. RT qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of any RNA template (mrna, total RNA, viral RNA), low copy number genes highqu 1Step RT qpcr mastermixes in combination with a blend of thermostable and extremely active Reverse Transcriptase & advanced RNase Inhibitor (RT Mix) allow for a single step one tube RT qpcr. qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT/GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. To suit the broad instrument range the 1Step RT qpcr Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 10. Reverse transcription & qpcr in one tube with highest sensitivity Efficient cdna synthesis ensured by the thermostable Reverse Transcriptase & advanced RNase Inhibitor blend Universal Kit for standard and fast cycling, GC/AT rich templates Rapid extension, early Ct Supplied with PCR Water for maximum convenience PRECAUTIONS FOR WORK WITH RNA Take care to prevent RNA from degradation by widely spread and stable RNases. Prepare crude samples and set up reactions in different dedicated areas, use DEPC-treated nuclease-free labware and gloves. Before the cdna synthesis, check RNA quality on denaturing agarose gel to be sure you have good quality material. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control, no RT Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Higher amounts of RT3 Mix improve Ct, but primer dimers may appear. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe 200 nm final c. (0.4 µl of 10 µm) Total RNA template or 1 pg to 1 µg or mrna template >0.01 pg PCR Water to 10 l 1Step RT qpcr Mix, 2X 10 l RT3 Mix, 20X 1-2 l Mix gently, avoid bubbles. Place into the instrument set like: Reverse Transcription 1 cycle: C - 10 min Initial denaturation 1 cycle: 95 C - 2 min Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. 12 Info Tel: info@highqu.com

15 1Step RT qpcr Probe ROX H Kit 2 x 1 ml - 1Step RT qpcr Probe ROX H Mix, 2X QOP r of 20 µl 1 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 2 x 1 ml - PCR Water 10 x 1 ml - 1Step RT qpcr Probe ROX H Mix, 2X QOP r of 20 µl 5 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 10 x 1 ml - PCR Water Storage In the dark at -20 C. Hot Start Taq, dntps at 0.25 mm, optimized buffer, high ROX concentration. 20X concentrated blend of modified MMuLV RT and RNase Inhibitor. qpcr on instruments calibrated with high ROX conc. RT qpcr assays based on specific probes: including TaqMan, Molecular Beacons, Scorpions Probes Quantification of any RNA template (mrna, total RNA, viral RNA), low copy number genes highqu 1Step RT qpcr mastermixes in combination with a blend of thermostable and extremely active Reverse Transcriptase & advanced RNase Inhibitor (RT Mix) allow for a single step one tube RT qpcr. qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum optimization. To suit the broad instrument range the 1Step RT qpcr Probe Mixes are available in three versions without ROX, with low or high ROX concentration. See the selection table on page 10. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control, no RT Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Higher amounts of RT3 Mix improve Ct, but primer dimers may appear. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Reverse transcription & qpcr in one tube with highest sensitivity Efficient cdna synthesis ensured by the thermostable Reverse Transcriptase & advanced RNase Inhibitor blend Universal Kit for standard and fast cycling, GC/AT rich templates Rapid extension, early Ct PCR Water supplied for maximum convenience PRECAUTIONS FOR WORK WITH RNA Take care to prevent RNA from degradation by widely spread and stable RNases. Prepare crude samples and set up reactions in different dedicated areas, use DEPC-treated nuclease-free labware and gloves. Before the cdna synthesis, check RNA quality on denaturing agarose gel to be sure you have good quality material. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Specific Probe 200 nm final c. (0.4 µl of 10 µm) Total RNA template or 1 pg to 1 µg or mrna template >0.01 pg PCR Water to 10 l 1Step RT qpcr Mix, 2X 10 l RT3 Mix, 20X 1-2 l Mix gently, avoid bubbles. Place into the instrument set like: Reverse Transcription 1 cycle: C - 10 min Initial denaturation 1 cycle: 95 C - 2 min Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. order@highqu.com Order Tel:

16 1Step RT qpcr Green ROX L Kit QOD r of 20 µl 2 x 1 ml - 1Step RT qpcr Green ROX L Mix, 2X 1 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 2 x 1 ml - PCR Water 10 x 1 ml - 1Step RT qpcr Green ROX L Mix, 2X QOD r of 20 µl 5 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 10 x 1 ml - PCR Water Storage In the dark at -20 C. Hot Start Taq, dntps at 0.25 mm, optimized buffer, low ROX concentration. 20X concentrated blend of modified MMuLV RT and RNase Inhibitor. Relative gene expression analysis, absolute quantification of any RNA template (mrna, total RNA, viral RNA), low copy number genes Intercalating dye fluorescence based qpcr on instruments calibrated with low ROX conc. highqu 1Step RT qpcr mastermixes in combination with a blend of thermostable and extremely active Reverse Transcriptase & advanced RNase Inhibitor (RT Mix) allow for a single step one tube RT qpcr. qpcr mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. To suit the broad instrument range the 1Step RT qpcr Green Mixes are available in different versions with low or high ROX concentration. See the selection table on page 10. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control, no RT Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Higher amounts of RT3 Mix improve Ct, but primer dimers may appear. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for this step. Reverse transcription & qpcr in one tube with highest sensitivity Efficient cdna synthesis ensured by the thermostable Reverse Transcriptase & advanced RNase Inhibitor blend Universal Kit for standard and fast cycling, GC/AT rich templates Rapid extension, early Ct PCR Water supplied for maximum convenience PRECAUTIONS FOR WORK WITH RNA Take care to prevent RNA from degradation by widely spread and stable RNases. Prepare crude samples and set up reactions in different dedicated areas, use DEPC-treated nuclease-free labware and gloves. Before the cdna synthesis, check RNA quality on denaturing agarose gel to be sure you have good quality material. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Total RNA template 1 pg to 1 µg or mrna template >0.01 pg PCR Water to 10 l 1Step RT qpcr Mix, 10 l RT3 Mix, 20X 1-2 l Mix gently, avoid bubbles. Place into the instrument set like: Reverse Transcriptio 1 cycle: C - 10 min Initial denaturation 1 cycle: 95 C - 2 min Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. 14 Info Tel: info@highqu.com

17 1Step RT qpcr Green ROX H Kit QOD r of 20 µl 2 x 1 ml - 1Step RT qpcr Green ROX H Mix, 2X 1 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 2 x 1 ml - PCR Water 10 x 1 ml - 1Step RT qpcr Green ROX H Mix, 2X QOD r of 20 µl 5 x 0.2 ml - RT3 Mix, 20X (RNase Inhibitor+RTase) 10 x 1 ml - PCR Water Storage In the dark at -20 C. Hot Start Taq, dntps at 0.25 mm, optimized buffer, high ROX concentration. 20X concentrated blend of modified MMuLV RT and RNase Inhibitor. Relative gene expression analysis, absolute quantification o Reverse transcription & qpcr in one tube with highest sensitivity any RNA template (mrna, total RNA, viral RNA), low copy Efficient cdna synthesis ensured by the thermostable Reverse number genes Transcriptase & advanced RNase Inhibitor blend Intercalating dye fluorescence based qpcr on instruments Universal Kit for standard and fast cycling, GC or AT rich templates calibrated with high ROX conc. Rapid extension, early Ct PCR Water supplied for maximum convenience PRECAUTIONS FOR WORK WITH RNA highqu 1Step RT qpcr mastermixes in combination with a Take care to prevent RNA from degradation by widely spread and blend of thermostable and extremely active Reverse stable RNases. Prepare crude samples and set up reactions in differe Transcriptase & advanced RNase Inhibitor (RT Mix) allow for dedicated areas, use DEPC-treated nuclease-free labware and gloves single step one tube RT qpcr. qpcr mastermixes are based on the small molecular inhibito Before the cdna synthesis, check RNA quality on denaturing agarose technology Hot Start PCR allowing to achieve highest sensitiv gel to be sure you have good quality material. and specificity under both standard and fast qpcr cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimization. To suit the broad instrument range the 1Step RT qpcr Green Mixes are available in different versions with low or high RO concentration. See the selection table on page 10. Use special primer selection programs for good planning. Work with amplicons in a range of , max 400 bp. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Run reactions in triplets; include a no-template control, no Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Higher amounts of RT3 Mix improve Ct, but primer dimers may appear. Do not perform annealing/extension for more than 30 seconds and do not use lower than 60 C temperature for step. Prepare a 20 µl reaction: Reverse Primer nm final c. Forward Primer nm final c. Total RNA template or 1 pg to 1 µg or mrna template >0.01 pg PCR Water to 10 l 1Step RT qpcr Mix, 2X 10 l RT3 Mix, 20X 1-2 l Mix gently, avoid bubbles. Place into the instrument set like: Reverse Transcription 1 cycle: C - 10 min Initial denaturation 1 cycle: 95 C - 2 min Denaturation 40 cycles: 95 C - 5 sec Annealing/extension 40 cycles: C sec Follow instrument instructions for melting curve analysis. For optional use, the ROX passive reference dye is premixed within the ROX L and ROX H qpcr Mixes. If the purchaser has an instrument capable of optional ROX detection and wishes to perform the optional normalization of the signal, then the user must select the option in the software. Notice to Purchaser: With purchasing of this product, no rights are conveyed with respect to U.S. Patent: 5,928,907 and corresponding patents outside the US. order@highqu.com Order Tel:

18 End-point PCR Selection: Standard, Hot-start, Long, High fidelity, Direct PCR Standard Hot Start Robust, Long & Hot Start High Fidelity DNA Polymerase Mastermix Mastermix Taq DNA Polymerase Start Taq Polymerase Start Taq Mastermix Polymerase Mastermix DNA Polymerase page 17 page 18 page 19 page 20 page 21 page 22 page 23 page 24 page 25 Fast cycling GC/AT rich PCR Hot Start High sensitivity Fidelity vs Taq 1 X 1 X 1 X 1 X 1 X 1 X 5 X 5 X 50 X Long PCR Max. amplicon 6 kb 6 kb 6 kb 5 kb 6 kb 6 kb 35 kb 35 kb 10 kb High yields Direct PCR Colony Colony Colony Colony Colony, blood, urine Colony, blood, urine Colony, blood Colony, blood Colony Multiplex PCR Classical PCR Cloning TA TA TA TA TA TA TA TA Blunt Direct Loading on gels * with dntps Mastermix * and magnesium concentrations, PCR enhancers and stabilizers what allows for easier amplification of GC or AT rich templates and for success in fast cycling. 16 Info Tel: info@highqu.com

19 Taq DNA Polymerase PCE0101 PCE0105 Storage 500 u - Taq DNA Polymerase, 5 u/µl 500 u 4 x 1 ml - 5 x 500 u u 20 x 1 ml - 5 In the dark at -20 C. Enzyme in storage buffer. 2, enhancers, stabilizers. Routine PCR up to 6 kb Amplification of complex (GC/AT rich) templates Colony PCR Fast PCR TA cloning highqu Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling. Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 10 4 (a number of correct nucleotides incorporated before the first error) and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the 2X Red Taq Mastermix and 2X Taq Mastermix are available. See next pages. Engineered Taq combined with advanced buffer - a synergy providing advantages over classical Taq Polymerases Higher yields under standard and fast cycling Increased success in amplification of longer templates (6 kb) Robust amplification of GC/AT rich templates 2+ and dntps PERFORMANCE highqu sensitivity compared to competitor Taq DNA Polymerase (below). PCR of a 1.2 kb fragment of 60% GC GAPDH, from human genomic DNA, in a 3 fold dilution from left to right. Starting from 200 ng of DNA up to 0.7 pg in the 7th dilution. Prepare a 50 µl reaction: Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng PCR Buffer 10 µl Water (PCR Water, to 49 l WAT0110) Taq DNA µl Polymerase, 5 u/µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C - 1 min Denaturation 40 cycles: 95 C - 15 sec Annealing 40 cycles: C 15 sec Extension 40 cycles: 72 C 1-90 sec (15 sec/kb) Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

20 - NEW 5 x 1 ml - PCM r of 50 µl 5 x 1 ml - PCR Water 25 x 1 ml - Red Taq Mastermix, 2X PCM r of 50 µl 25 x 1 ml - PCR Water Storage In the dark at -20 C. 1X mastermix contains 0.25 mm dntps, 3 mm MgCl 2, enhancers, stabilizers, red electrophoresis tracking dye and density reagents for gel loading Routine PCR up to 6 kb with a direct gel loading option Amplification of complex (GC/AT rich) templates Colony PCR Fast PCR TA cloning Engineered Taq combined with advanced buffer - a synergy providing advantages over classical Taq Polymerases Premixed with red dye and density reagents for direct loading on the gels after the PCR Higher yields under standard and fast cycling Increased success in amplification of longer templates (6 kb) Robust amplification of GC/AT rich templates highqu Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling. Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 10 4 (a number of correct nucleotides incorporated before the first error) and produces A-tailed products suitable for ligating into TA cloning vectors. The convenience of the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. density reagents for direct loading on the gels after the PCR. In a 2% agarose TAE gel the dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments. The mastermix is even supplied with PCR water, and the only thing to add is the template. Prepare a 50 µl reaction: Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. dye and density reagents for direct loading on the gels after the PCR. In a 2% agarose TAE gel the dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments. Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng PCR Water to 25 l Red Taq Mastermix, 2X 25 µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C - 1 min Denaturation 40 cycles: 95 C - 15 sec Annealing 40 cycles: C 15 sec Extension 40 cycles: 72 C 1-90 sec (15 sec/kb) Load probes on the agarose gel. The red loading dye is included in the mastermix. Store probes for short time on ice, for long at -20 C. 18 Info Tel: info@highqu.com

21 PCM r of 50 µl 5 x 1 ml - 5 x 1 ml - PCR Water 1X mastermix contains 0.25 mm dntps, 3 mm MgCl 2, enhancers, PCM r of 50 µl 25 x 1 ml - stabilizers. 25 x 1 ml - PCR Water Storage In the dark at -20 C. Routine PCR up to 6 kb Amplification of complex (GC/AT rich) templates Colony PCR Fast PCR TA cloning Engineered Taq combined with advanced buffer - a synergy providing advantages over classical Taq Polymerases Higher yields under standard and fast cycling Increased success in amplification of longer templates (6 kb) Robust amplification of GC/AT rich templates highqu Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling. Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 10 4 (a number of correct nucleotides incorporated before the first error) and produces A-tailed products suitable for ligating into TA cloning vectors. The convenience of Taq DNA Polymerase is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR water, and the only thing to add is the template. Prepare a 50 µl reaction: Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng PCR Water to 25 l Taq Mastermix, 2X 25 µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C - 1 min Denaturation 40 cycles: 95 C - 15 sec Annealing 40 cycles: C 15 sec Extension 40 cycles: 72 C 1-90 sec (15 sec/kb) Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

22 Taq DNA Polymerase 1500 u - Taq DNA Polymerase, 5 u/µl PCE u 4 x 2 ml - 10X PCR Buffer Enzyme in storage buffer. PCE u 2 x 2 ml - 50 mm MgCl 2 2 x 1500 u - Taq DNA Polymerase, 5 u/µl 8 x 2 ml - 10X PCR Buffer 10X PCR Buffer contains enhancers and stabilizers, but no dntps and no Mg x 2 ml - 50 mm MgCl 2 Storage In the dark at -20 C. Routine PCR up to 5 kb RT-PCR, Colony PCR TA cloning, library construction Genotyping, screening highqu Taq DNA Polymerase is the classical enzyme for routine PCR applications providing high amplification yields of 3-5 kb targets under various conditions. Taq DNA Polymerase is purified from a recombinant E. coli strain carrying the Taq DNA polymerase gene. Taq DNA polymerase is thermostable 5 3 DNA polymerase. It lacks 3 5 exonuclease (proofreading) activity and has low 5 3 exonuclease activity. Polymerase exhibits deoxynucleotidyl transferase activity resulting in A-overhang at the 3'-ends of PCR products and allowing for TA cloning. The PCR accuracy of Taq DNA Polymerase is 4.5 x 10 4 (a number of nucleotides incorporated before the first error occurs). High yields in routine PCR, good performance in fast PCR Guarantied successful DNA preparation for TA cloning Robust on complex templates RECOMMENDATIONS The supplied reaction buffer contains no dntps and the 50 mm MgCl 2 is provided separately what allows for magnesium optimization upon the need. dntps in mixes or sets can be purchased separately (page 28) Typical concentration of each dntp in the reaction is mm. Higher concentration increase yields, however Mg 2+ -ions bind to dntps, therefore, both components shall be present in coordinated concentrations. Too high dntps and magnesium concentrations reduce PCR fidelity. Mix well each dntp and magnesium solution. Use final 3 mm MgCl 2 and 0.25 mm each dntp concentrations for routine PCR. Starting dntp Mix conc. Vol. of dntp mix in 50 µl r. Final Mg 2+ conc. in r. 10 mm 1.25 µl 2 mm 2 µl 25 mm 0.5 µl 3 mm 3 µl Vol. of 50 mm MgCl 2 in 50 µl rxn to achieve desired conc. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time. Taq DNA Polymerase speed is ~ 2000 nucleotides/min, so seconds of extension can be provided per cycle, depending on amplicon size. Start annealing with 55 C and perform gradient by increasing temperature in 2 C up to 65 C to choose the best. Calculate primer annealing temperature using software. Prepare a 50 µl reaction: Rev. & For. Primers µm final (1-2 µl of 10 µm each) cdna Template or <100 ng or gdna Template ng 10X PCR Buffer 5 µl dntp Mix 0.25 mm final (1.25 µl of 10 mm dntp mix) (NUM0201) 50 mm Mg Cl 2 3 l Water (PCR Water, to 49 l WAT0110) Taq DNA µl Polymerase, 5 u/µl Mix gently, avoid bubbles. Place into the instrument set: Initial denaturation 1 cycle: 95 C 60 sec Denaturation cycles: 95 C - 15 sec Annealing cycles: C 15 sec Extension cycles: 72 C sec Final extension 1 cycle: 72 C 5 min (for TA cloning) Store probes for short time on ice, for long at -20 C. 20 Info Tel: info@highqu.com

23 Hot Start Taq Polymerase HSE0101 HSE0105 Storage 2x 250 u - Taq Polymerase, 5 u/µl 500 u 4 x 1 ml - 10 x 250 u - Polymerase, 5 u/µl 2500 u 20 x 1 ml - In the dark at -20 C. Enzyme in storage buffer. MgCl 2, enhancers, stabilizers. Sensitive hot-start PCR up to 6 kb Low copy target detection Amplification of complex (GC/AT rich) templates Fast PCR TA cloning Multiplex hot-start PCR Small molecular inhibition hot-start technology combined with advanced buffer - advantages over classical hot- start Taq Outperforming sensitivity & specificity - low copy target detection Higher yields under standard and fast cycling Increased success in amplification of longer templates (6 kb) Robust amplification of GC rich templates Mg 2+ and dntps highqu Hot Start Taq Polymerase is the superior sensitive hot-start enzyme. The activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background. In combination with the optimized ALLin buffer enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling. Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the 2X Hot Start Taq Mastermix is available. See next page. PERFORMANCE Start Taq Competitor K Competitor L Start Taq DNA Polymerase shows better yields and higher sensitivity compared to competitors. PCR of a 0.4 kb fragment, from human genomic DNA, under fast cycling conditions. Starting template 100 ng with further 10 fold dilutions. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Prepare a 50 µl reaction: Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng 10 µl Water (PCR Water, WAT0110) to 49 l Hot Start Taq µl DNA Polymerase, 5 u/µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C 1-2 min Denaturation 40 cycles: 95 C - 15 sec Annealing 40 cycles: C 15 sec Extension 40 cycles: 72 C 1-90 sec (15 sec/kb) Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

24 Hot Start Taq Mastermix, 2X HSM r of 50 µl 5 x 1 ml - Mastermix, 2X 5 x 1 ml - PCR Water 1X mastermix contains 0.25 mm dntps, 3 mm MgCl 2, HSM r of 50 µl 25 x 1 ml - enhancers, stabilizers. 25 x 1 ml - PCR Water Storage In the dark at -20 C. Sensitive hot-start PCR up to 6 kb Low copy target detection Amplification of complex (GC/AT rich) templates Fast PCR TA cloning Multiplex hot-start PCR Small molecular inhibition hot-start technology combined with advanced buffer - advantages over classical hot- start Taq Outperforming sensitivity & specificity - low copy number target detection and no background Higher yields under standard and fast cycling Increased sensitivity and success in amplification of longer templates (6 kb) Robust amplification under difficult conditions - GC rich templates highqu Hot Start Taq DNA Polymerase is the superior sensitive enzyme The activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background. In combination with the optimized ALLin buffer enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling. Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, and produces A-tailed products suitable for ligating into TA cloning vectors. The convenience of Hot Start Taq DNA Polymerase is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR water, and the only thing to add is the template. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <6kb and 1 sec/kb - <1kb. Use >90 sec extension for multiplexing. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Prepare a 50 µl reaction: Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng PCR Water to 25 l Hot Start Taq Mastermix, 2X 25 µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C 1-2 min Denaturation 40 cycles: 95 C - 15 sec Annealing 40 cycles: C 15 sec Extension 40 cycles: 72 C 1-90 sec (15 sec/kb) Store probes for short time on ice, for long at -20 C. 22 Info Tel: info@highqu.com

25 HLE0101 HLE0105 Storage 250 u u 2 x 1 ml - 5 x 250 u u 10 x 1 ml - In the dark at -20 C. Enzyme in storage buffer. 2, enhancers, stabilizers. Amplification of difficult & complex (GC/AT rich) templates Long PCR (up to 35 kb) with higher fidelity Colony & crude sample PCR Multiplex PCR TA cloning RPH Robust, Proofreading, Hot-start Polymerase Low-copy number target detection ensured by small molecular inhibitor hot-start Long PCR (up to 35 kb), higher-fidelity (5X higher than Taq) ensured by proofreading activity High yields under standard and fast cycling Robust - GC or AT rich templates, crude sample PCR 2+ and dntps highqu Polymerase (Robust, Proofreading, Hotstart Polymerase) is the versatile engineered enzyme combining best polymerase properties for excellence in most demanding PCR applications, like low copy detection, long or high fidelity PCR, amplification of complex templates, crude sample PCR and multiplexing. has 5 times higher fidelity than Taq DNA Polymerase and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the 2X Mastermix is available. See next page. PERFORMANCE Competitor R Competitor L higest sensitivity amplification of 25 kb target from lowest amounts of human genomic DNA. The starting template concentration is 200 ng of human genomic DNA with 2x further dilutions. 25kb fragment of the p53 gene was amplified. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <5kb and 40 to 60 sec/kb - <5kb-35 kb. Use >90 sec extension for multiplexing. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Prepare a 50 µl reaction: Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng 10 µl Water (PCR Water, to 49 l WAT0110) µl Polymerase, 5 u/µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C - 1 min Denaturation cycles: 95 C - 15 sec Annealing cycles: C 15 sec Extension cycles: 72 C 10 min Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

26 HLM r of 50 µl 5 x 1 ml - 5 x 1 ml - PCR Water 1X mastermix contains 0.25 mm dntps, 3 mm MgCl 2, HLM r of 50 µl 25 x 1 ml - enhancers, stabilizers. 25 x 1 ml - PCR Water Storage In the dark at -20 C. Amplification of difficult & complex (GC/AT rich) templates Long PCR (up to 35 kb) with higher fidelity Colony & crude sample PCR Multiplex PCR TA cloning RPH Robust, Proofreading, Hot-start Polymerase Low-copy number target detection ensured by small molecular inhibitor hot-start Long PCR (up to 35 kb), higher-fidelity (5X higher than Taq) ensured by proofreading activity High yields under standard and fast cycling Robust - GC or AT rich templates, crude sample PCR highqu Robust, Proofreading, Hotstart Polymerase) is the versatile engineered enzyme combining best polymerase properties for excellence in most demanding PCR applications, like low copy detection, long or high fidelity PCR, amplification of complex templates, crude sample PCR and multiplexing. olymerase has 5 times higher fidelity than Taq DNA Polymerase and produces A-tailed products suitable for ligating into TA cloning vectors. The convenience of is maximized by the use of 2X Mastermix providing the additional advantage of reduced pipetting and minimized errors. The mastermix is even supplied with PCR water, and the only thing to add is the template. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 15 sec/kb - <5kb and 40 to 60 sec/kb - <5kb-35 kb. Use >90 sec extension for multiplexing. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Do not use fast cycling for multiplexing. Prepare a 50 µl reaction: Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng PCR Water to 25 l Mastermix, 2X 25 µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C 1-2 min Denaturation cycles: 95 C - 15 sec Annealing cycles: C 15 sec Extension cycles: 72 C 10 min Store probes for short time on ice, for long at -20 C. 24 Info Tel: info@highqu.com

27 HLE0201 HLE0205 Storage 200 u - DNA Polymerase, 2 u/µl 200 u 3 x 1 ml - 5 x 200 u u 15 x 1 ml - In the dark at -20 C. Enzyme in storage buffer. MgCl 2, enhancers, stabilizers. High-fidelity PCR up to 10 kb Long PCR up to 10 kb Amplification of complex (GC/AT rich) templates Fast high-fidelity PCR Blunt cloning Crude sample PCR with high fidelity highqu HiFi DNA Polymerase is the outperforming high-fidelity enzyme derived from Pfu polymerase by introducing several point mutations. The robust engineered enzyme in combination with the optimized ALLin buffer provides higher fidelity (50X higher than Taq), better performance, increased success in demanding applications like amplification of complex or longer templates, crude sample PCR and fast cycling. HiFi DNA Polymerase produces blunt-ended products suitable for ligating into blunt vectors. Foe endpolishing and phosphorylation of DNA ends before bluntcloning use the HighEnd Repair Kit (HER0101). Engineered proofreading enzyme (Pfu-based) and advanced buffer 50 x higher fidelity than classic Taq Increased sensitivity and high yields under standard and fast cycling Increased success in PCR of longer templates (10 kb) Robust on GC/AT rich templates, crude samples Mg2+ and dntps PERFORMANCE highqu HiFi DNA Polymerase (above) shows better yields and higher sensitivity compared to P enzyme from competitor N (below). PCR of a 1 kb fragment of 60% GC GAPDH, from human genomic DNA. The template is diluted 2 fold over 8 orders of magnitude, starting from 100 ng. Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. The longer the amplicon, the longer the extension time: 30 sec/kb. Run an annealing temperature gradient from 55 C to 65 C to choose the best specificity conditions. Prepare a 50 µl reaction: Rev. & For. Primers µm final each ( 2 µl of 10 µm) cdna Template or <100 ng or gdna Template ng HiFi Buffer 10 µl Water (PCR Water, to 49 l WAT0110) HiFi DNA Polymerase, 2 u/µl 0.5 µl Mix gently, avoid bubbles. Place into the instrument set like: Initial denaturation 1 cycle: 95 C 1 min Denaturation cycles: 95 C - 15 sec Annealing cycles: C 15 sec Extension cycles: 72 C 30 sec (30 sec/kb) Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

28 RT PCR & Reverse Transcription Selection 1Step RT PCR Kit Transcriptase Synthesis Kit SecurRIN RNase Inhibitor page 27 page 28 page 29 page 30 Short description One-step RT PCR in one tube cdna synthesis of long transcripts at elevated temperature Unbiased cdna synthesis for qpcr Advanced RNA protection from RNases Enhanced cdna synthesis NA GC rich and complex templates NA Full-length cdna transcripts up to 15 kb NA High sensitivity NA One-step RT-PCR NA Two-step RT-PCR NA Two-step RT-qPCR NA RNA protection from RNases NA Not Applicable 26 Info Tel:

29 1Step RT PCR Kit RTK0201 Storage 2 x 1.25 ml - 1Step RT PCR Mastermix, 2X 100 r of 2 x ml - RT2 Mix, 20X 50 µl 3 x 1 ml - PCR Water In the dark at -20 C. 1X mastermix contains hot-start Taq DNA Polymerase, 0.25 mm dntps, 3 mm MgCl 2, enhancers, stabilizers. RT2 Mix is a 20X concentrated blend of reverse transcriptase and RNase inhibitor. One step RT-PCR RT-PCR of complex GC/AT rich templates Fast RT-PCR TA cloning Easy to use combination of the RT mix with the RT-PCR mastermix - reverse transcription and PCR in one tube RT Mix contains RNase inhibitor and thermostable reverse transcriptase (up to 55 C) allowing for high cdna yields RT PCR Mastermix allows for sensitive low copy number targets detection due to proprietary hot-start High yields under standard and fast cycling conditions and on GC/AT rich templates highqu 1Step RT PCR Kit combines the blend of Reverse Transcriptase and RNase Inhibitor for efficient reverse transcription and the PCR Mastermix for subsequent amplification of cdna in the same tube. RT2 Mix, 20X is a blend of the engineered MMuLV (stable at C allowing for high yields of long transcripts) with an efficient Ribonuclease Inhibitor protecting the template RNA from RNases. The resulting cdna is then amplified by the 1Step RT PCR Mastermix, 2X. The PCR mastermix contains our proprietary Hot Start Taq DNA Polymerase. The activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background. RNA is extremely sensitive to degradation by RNases present everywhere. Take care to protect RNA from degradation and to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include a no-template control, no RT2 Mix control and positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Perform cdna synthesis 10 min at 45 C, 20 min for >1 kb, increase temperature to 55 C for complex templates. The longer the amplicon, the longer the extension: use 15 sec/kb for < 3 kb targets, increase time for >3 kb. Run an annealing temperature gradient from 58 C to 65 C to choose the best specificity conditions. In combination with the optimized buffer the enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling. Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the Kit includes even the PCR Water to set up the reaction, so the only thing you need to take care is the high quality RNA template. Prepare a 50 µl reaction: Rev. & For. Primers µm final (2 µl of 10 µm each) Total RNA or 1 pg to 1 µg or mrna >0.01 pg PCR Water to 22.5 µl 1Step RT PCR 25 µl Mastermix, 2X RT2 Mix, 20X 2.5 µl Mix gently, avoid bubbles. Place into the instrument set like: Reverse 1 cycle: C 10 to 20 min transcription Initial denaturation 1 cycle: 95 C 2 min Denaturation 40 cycles: 95 C - 10 sec Annealing 40 cycles: C 10 sec Extension 40 cycles: 72 C sec (15 sec/kb) Store probes for short time on ice, for long at -20 C. order@highqu.com Order Tel:

30 Reverse Transcriptase - NEW RTE0301 RTE0305 Storage 50 µl - Reverse Transcriptase, 200 u/µl u 0.5 ml - 5X Reaction Buffer 5 x 50 µl - Reverse Transcriptase, 200 u/µl u 5 x 0.5 ml - 5X Reaction Buffer In the dark at -20 C. Storage buffer contains Tris, 50% glycerol and other components. 5X Reaction Buffer contains Tris, MgCl 2, DTT and other components. cdna synthesis of up to 15 kb long transcripts Template generation for RT-PCR & RT-qPCR cdna synthesis from complex templates Thermostable Reverse Transcriptase for efficient cdna synthesis High yields of full lengths transcripts up to kb cdna synthesis from complex templates at up to 55 C High sensitivity detection from 1 pg of total RNA template The HighScriber Reverse Transcriptase is a premium tool for the high efficiency reverse transcription of up to kb long cdna. HighScriber Reverse Transcriptase allows for high detection sensitivity from 1 pg of total RNA. The wide reaction temperature range (38 C - 55 C) ensures efficient cdna synthesis from complex or GC rich templates. The enzyme uses ssrna or ssdna as a template, possesses no detectable Ribonuclease H activity specific to RNA in RNA-DNA hybrids. A highly reduced Ribonuclease H activity allows for transcription of full lengths long transcripts. HighScriber Reverse Transcriptase can be used for RACE as it has terminal transferase activity - cdna. RNA is extremely sensitive to degradation by RNases present everywhere. Take care to protect RNA from degradation keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Optionally, add SecurRIN RNase Inhibitor (RNI0201) in to the cdna synthesis reaction (~20 units for 20 µl reaction). Check the integrity of RNA prior to cdna synthesis in denaturing agarose gel. Include positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Use final mm dntp concentration for cdna synthesis. For best results, optimize the template and primer amount. Do not exceed the recommended amount of the enzyme. Perform reaction for min, for short transcripts min are sufficient. Choose optimal reaction temperature in a range of C. PRODUCT SPECIFICATIONS Optimal activity at C Temperature range C Inactivation at 85 C for 10 min Unit Definition One unit is defined as the amount of enzyme that will incorporate 1 nmol of dttp into acid-insoluble material in a total reaction volume of 50 l in 10 minutes at 37 C using poly (ra) oligo (dt) 18 as template. Prepare a 20 µl reaction: SecurRIN RNase Inhibitor 0. 5 µl (20 u) (optional), 40 u/µl 10 mm dntp Mix (NUM0201) 2 µl (0.8-1 mm final) Total RNA or 1 pg to 5 µg or Poly-A mrna 1 pg to 0.5 µg Oligo dt primer or 0.5 µg or Random primer or 0.2 µg or Specific primer pmol Water (PCR Water, WAT0110) to 15 µl Mix gently, avoid bubbles. Heat 5 min at 65 C, spin, place on ice for 1 min. Add the 4 µl of 5X HighScriber Buffer Add 1 µl HighScriber RTase, 200 u/µl and mix well. Incubate 2 min at 42 C for Oligo dt and for Specific primer or 10 min at 25 C for Random primer to anneal. Incubate min at 50 C to synthesize cdna. Inactivate at 85 C for 10 min. Store reactions at -20 C or on ice for an immediate use. Use 2-5 µl of this reaction mix per 50 µl PCR reaction. Use 1-2 µl of this reaction mix per 20 µl qpcr reaction. 28 Info Tel: info@highqu.com

31 qcdna Synthesis Kit - NEW 25 µl - Enzyme Blend, 20X RTK r of 20 µl 100 µl - Reaction Mix 1 ml PCR Water 4 x 25 µl - Enzyme Blend, 20X RTK r of 20 µl 4 x 100 µl - Reaction Mix 2 x 1 ml PCR Water Storage In the dark at -20 C. cdna template generation for qpcr or PCR Unbiased, efficient cdna synthesis Detection of low target amounts cdna synthesis from complex templates The qscriber cdna Synthesis Kit is a highly efficient and simple-to-use system for cdna synthesis eliminating the need for tedious reaction optimization. The Enzyme Blend ensures high sensitivity detection from low copy number targets. The highly active and thermostable HighScriber Reverse Transcriptase blended with RNase Inhibitor allows for an efficient cdna synthesis and reaction safety. The wide reaction temperature range (38 C - 55 C) ensures efficient transcription from GC rich templates. The 5X q Reaction Mix includes optimal concentrations of magnesium and dntps and a combination of anchored oligo (dt) and random hexamers for unbiased representation of mrna ends. The kit is an optimal choice for generating high quality cdna from viral RNA, mirna or other targets for qpcr or for PCR. Enzyme Blend Storage buffer contains Tris, 50% glycerol and other components. Reaction Mix contains dntps, MgCl 2, anchored oligo(dt), random hexamers and other components. with Ribonuclease Inhibitor for efficient cdna synthesis Optimized reaction mix with oligo (dt) and random primers for unbiased representation of mrna ends cdna synthesis from complex templates at up to 55 C High sensitivity detection from 1 pg of total RNA template PERFORMANCE q provides excellent results within the very broad range of the total RNA amount used 4 pg, 40 pg, 400 pg, 4 ng, 40 ng, 400 ng and 4 µg of mouse total RNA were used for cdna synthesis under the standard q protocol conditions. An aliquot from each reaction was taken for subsequent qpcr with lify a 70 bp fragment of the mouse RN18S gene. All reactions independently from the initial amount of RNA were 100% efficient. Prepare a 20 µl reaction: RNA is sensitive to degradation by RNases present everywhere. Take care to protect RNA from degradation keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Include positive and negative controls in parallel. Thaw and keep reagents on ice. Mix well before use. Do not add any other components into the reaction with exception of the template and the supplied reagents. The recommended reaction temperature is C. For GC rich templates, the temperature can be increased up to 55 C. 5X Reaction Mix 4 µl Enzyme Blend, 20X 1 µl Total RNA 1 pg to 5 µg PCR Water Up to 20 µl Mix gently, avoid bubbles. Incubate 30 min at C to synthesize cdna. Inactivate the enzyme at 85 C for 10 min. Store reactions at -20 C or on ice for an immediate use. Use 2-4 µl of this reaction mix per 20 µl qpcr reaction. order@highqu.com Order Tel:

32 RNase Inhibitor - NEW RNI u 75 µl - SecurRIN RNase Inhibitor, 40 u/µl Storage buffer contains 50% glycerol and other RNI u 500 µl - SecurRIN RNase Inhibitor, 40 u/µl components. Storage In the dark at -20 C Prevention of RNA degradation during: cdna synthesis RNA purification in vitro transcription The SecurRIN RNase Inhibitor is a premium tool for protection of the RNA from degradation during enzymatic reactions, storage or RNA purification. It is a recombinant protein derived from E. coli strain carrying the porcine RNase Inhibitor gene. SecurRIN is a 52 kda non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, RNase B, and RNase C. Efficient RNA protection from RNases during cdna synthesis or other procedures Economical robust high quality enzyme Active under different reaction conditions in different buffers Active in all common buffers used for RNA work 1 unit of the RNase Inhibitor is typically used for 1 microliter of RNA reaction UNIT DEFINITION One unit is defined as the amount of enzyme required to -cyclic monophosphate by 5 ng of RNAse A. example with HighScriber Reverse Transcriptase (RTE0301) RNA is extremely sensitive to degradation by RNases present everywhere. Take care to protect RNA from degradation keep your bench clean, wear gloves, use sterile tubes and filter pipet tips. Add SecurRIN RNase Inhibitor in to the cdna synthesis reaction (~20 units for 20 µl reaction). Check the integrity of RNA prior to cdna synthesis in denaturing agarose gel. Include positive control in parallel. Thaw and keep reagents on ice. Mix well before use. Use final mm dntp concentration for cdna synthesis. For best results, optimize the template and primer amount. Do not exceed the recommended amount of the enzyme. Perform reaction for min, for short transcripts min are enough. Choose optimal reaction temperature in a range of C. Prepare a 20 µl reaction: SecurRIN RNase Inhibitor, 40 u/µl 0. 5 µl (20 u) 10 mm dntp Mix (NUM0201) 2 µl (1 mm final) Total RNA or 1 pg to 5 µg or Poly-A mrna 1 pg to 0.5 µg Oligo dt primer or 0.5 µg or Random primer or 0.2 µg or Specific primer pmol Water (PCR Water, WAT0110) to 15 µl Mix gently, avoid bubbles. Heat 5 min at 65 C, spin, place on ice for 1 min. Add the 4 µl of 5X HighScriber Buffer Add 1 µl HighScriber RTase, 200 u/µl and mix well. Incubate 2 min at 42 C for Oligo dt and for Specific primer or 10 min at 25 C for Random primer to anneal. Incubate min at 50 C to synthesize cdna. Inactivate at 85 C for 10 min. Store reactions at -20 C or on ice for an immediate use. Use 2-5 µl of this reaction mix per 50 µl PCR reaction. Use 1-2 µl of this reaction mix per 20 µl qpcr reaction. 30 Info Tel: info@highqu.com

33 dntp Sets & Mixes dntp Mixes: NUM ml 1 ml - 25 mm dntp Mix Aqueous solution of equal concentration of 25 mm each of 4 dntps, ph 7.0 NUM ml 1 ml - 10 mm dntp Mix Aqueous solution of equal concentration of 10 mm each of 4 dntps, ph 7.0 dntp Sets: 0.25 ml -100 mm datp Aqueous solution of 100 mm datp, ph 7.0 NUS x 0.25 ml 0.25 ml mm dctp Aqueous solution of 100 mm dctp, ph ml mm dgtp Aqueous solution of 100 mm dgtp, ph ml mm dttp Aqueous solution of 100 mm dttp, ph 7.0 Storage In the dark at -20 C. All molecular biology applications including dntps, like: cdna synthesis Standard PCR, Long and high-fidelity PCR qpcr Sequencing Highest quality, >99% HPLC pure dntps for high & reproducible yields Pure from DNA contamination and from PCR inhibitors Highly stable remain pure after weeks at room temperature, after 30 freezing thawing cycles and during the 40 PCR cycles Available in ready-to use mixes and sets for maximized flexibility highqu dntp sets and mixes meet all highest industry standards and allow for unrivaled performance of your PCR and other DNA synthesis reactions. Produced under the stringent quality monitoring conditions, they guaranty reproducible results. More than 99% HPLC purity eliminates inhibitions of PCR and allows for increased yields with higher dntp concentrations. Exceptional stability eliminates dntp usability concerns related to short term ambient temperature shipments, room temperature storage or PCR exceeding 40 cycles. PREPARATION OF DNTP MIXES FROM A SET Highly concentrated solutions require thorough mixing before the use. The optimal dntp mix shall have equal concentrations of all 4 dntps. To prepare from a set of 4 dntps mixes of common concentrations, follow the guidelines below: Use same volume of each from PCR Resulting 1 ml Mix four 100 mm dntp solutions: Water concentration: 20 µl 920 µl 2 mm dntp 25 µl 900 µl 2.5 mm dntp 100 µl 600 µl 10 mm dntp 250 µl - 25 mm dntp RECOMMENDATIONS FOR STANDARD PCR Typical concentration of each dntp in the reaction is mm. Higher concentration increase yields, however Mg 2+ ions bind to dntps, therefore, both components shall be present in coordinated concentrations. Too high dntps and magnesium concentrations reduce PCR fidelity. Mix well each dntp and magnesium solution, to avoid concentration fluctuations. Use final 3 mm MgCl 2 with 0.25 mm each dntp concentration for routine PCR. Starting dntp Mix conc. Vol. of dntp mix for 50 µl rxn Final Mg 2+ conc. in rxn Vol. of 50 mm MgCl 2 for 50 µl rxn to achieve desired conc. 10 mm 1.25 µl 2 mm 2 µl 25 mm 0.5 µl 3 mm 3 µl order@highqu.com Order Tel:

34 PCR Water WAT x 1 ml 10 x 1 ml - PCR Water PCR-grade, nuclease free water Storage At -20 C. To minimize the effects of the contamination during the use it is not recommended to store water at room temperature. All molecular biology applications Pure PCR-grade, nuclease free water for excellence in molecular biology applications Same PCR Water is supplied with most of highqu products for maximum convenience and guaranteed performance Tested in most demanding PCR and qpcr applications highqu PCR Water is a supplementary high quality reagent for all demanding applications. It saves time being on your bench and guaranties purity of reactions and inhibitionfree performance of PCR reagents. highqu PCR Water is a deionized, membrane filtered water continuously tested in ultrasensitive qpcr and PCR applications, in amplification of long targets and highly specific detection of few copies of templates. Use PCR Water in all PCR applications like described in typical protocols. 32 Info Tel: info@highqu.com

35 - NEW RLK r of 20 µl 40 µl Rally T4 DNA Ligase, 1 r/µl 1.5 ml - 2X Rally Buffer 5 x 40 µl Rally T4 DNA Ligase, 1 r/µl RLK r of 20 µl 5 x 1.5 ml - 2X RallyBuffer Storage In the dark at -20 C. Storage buffer contains Tris, 50% glycerol and other components. 2X RallyBuffer contains DTT, ATP, PEG 6000 and other components. Cloning Vector-insert ligation Adaptor or linker ligation Linear DNA self-circularization Rally Rapid Ligation Kit is a premium tool designed for fast and highly efficient ligation reactions, cloning or adaptor/linker joining applications. The Kit contains Rally T4 DNA Ligase specially formulated to perform faster; and a 2X buffer which includes PEG to accelerate joining of DNA ends. The combination of both components allows for an efficient and fast ligation reaction of both blunt and cohesive DNA termini eliminating the need of hours or overnight incubations. Check the integrity and the concentration of the DNA prior the ligation. Include ligation positive control in parallel. Thaw and keep reagents on ice. Mix well before use. If precipitation in the buffer appears, warm it to dissolve. Use an insert : vector molar ratio around 3 : 1 to 6 : 1, optimize it if possible. Too much insert causes ligation of multiple inserts, too little reduces ligation efficiency. The Rally Buffer includes PEG which has almost no effect on transformation efficiency of chemicallycompetent cells, but reduces the transformation efficiency of electro-competent cells. Therefore, for electrotransformation PEG has to be removed from the mixture or highly diluted. Use only high efficiency competent cells for cloning. Check the transformation efficiency of the competent cells by transforming 0.1 ng of supercoiled vector DNA. Getting 100 colonies in this case means you have 1x10 6 transformants/1 µg supercoiled DNA. Take into account that the ligated mixture gives normally at least 10 times less transformants compared to the same amount of supercoiled DNA. 5 minutes fast high efficiency ligation Universal for both blunt or cohesive-end ligations Premium reagents - reproducible results PRODUCT SPECIFICATIONS Optimum activity at room temperature, around 25 C Inactivation at 65 C for 15 min Rally T4 DNA Ligase, same as the classical enzyme catalyzes the formation of a phosphodiester bond between phosphate and It joins both blunt and cohesive ends and repairs single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids. Prepare a 20 µl reaction: 2X Rally Buffer 10 µl Linear dephosphorylated vector DNA ~100 ng ( ng range) 100 ng of linear puc vectors (~2.7 kb) have ~0.1 pmol ends 100 ng of linear pbr322 vectors (~4.4 kb) have ~0.07 pmol ends Insert DNA ~200 ng (up to 3-6 X more (phosphorylated) pmol ends than vector) 200 ng of 1 kb linear DNA has ~0.6 pmol ends 100 ng of 0.5 kb linear DNA has ~0.6 pmol ends 40 ng of 0.2 kb linear DNA has ~0.6 pmol ends DNase-free water to 19 µl (PCR Water, WAT0110) Rally T4 DNA Ligase, 1 r/µl 1 µl maximum Mix well, incubate for 5-10 min at 25 C (room temperature). Use directly 1-5 µl of the mixture to transform 50 µl of chemically-competent cells. For electrotransformation PEG has to be removed from the mixture or diluted as follows: Option 1: re-purify the ligation mixture using PCR clean-up spin column kit, elute in 20 µl of water or TE and transform 1-2 µl of the eluate into 50 µl of electro-competent cells. Option 2: immediately after ligation dilute the mixture 10X by adding 180 µl of water or TE and transform 2-5 µl of the diluted mix into 100 µl of electro-competent cells. order@highqu.com Order Tel:

36 - NEW 40 µl HighEnd Repair Blend, 1 r/µl HER r of 25 µl 1.5 ml - 10X HighEndBuffer 0.5 ml - 1 mm dntp Mix 5 x 40 µl HighEnd Repair Blend, 1 r/µl HER r of 25 µl 2 x 1.5 ml - 10X HighEnd Buffer 3 x 0.5 ml - 1 mm dntp Mix Storage In the dark at -20 C. Enzyme blend storage buffer contains Tris, 50% glycerol and other components. 10X HighEndBuffer contains Tris, NaCl, MgCl 2, DTT, and other components. dntps serve as building blocks for filling-in reaction and as phosphate donors for phosphorylation. Preparation of PCR products, shared or nebulized DNA, restriction-digested DNA, cdna for blunt-end ligation - -phosphorylated blunt-ended ones HighEnd Repair Kit is a premium tool designed for rapid and highly efficient DNA end-repair before the ligation reactions. PCR products, shared or nebulized DNA, restriction-digested DNA and cdna can be blunted/phosphorylated in a couple of minutes and are ready for an efficient blunt-end ligation and cloning. The Kit includes HighEnd Repair Blend an optimized mix of T4 DNA Polymerase and T4 Polynucleotide Kinase. 3 polymerase and 35 exonuclease activities of T4 DNA Polymerase form the blunt-ended DNA. T4 Polynucleotide Kinase The resulting DNA is a high quality blunt-ended substrate for T4 DNA Ligase. Check the integrity and the concentration of the DNA prior the reaction. Always repurify PCR products before end-repair. Thaw and keep reagents on ice. Mix all components well before use. The optimal DNA amount in the reaction depends on the lengths of the DNA fragment. For example 1 µg of 1 kb linear DNA has ~3 pmol ends, but 1 µg of 100 bp linear DNA has 10 times more substrate for blunting/phosphorylation; i.e. even 30 pmol DNA ends. Therefore, the shorter is the DNA fragment, the less of it shall be used in micrograms for end-repair or for later ligation reaction. For high DNA amounts upscale the reaction accordingly. For example 5 µg of short 100 bp fragment can be endrepaired in 100 µl reaction using 2-5 µl of HighEnd Repair Blend. Fast and simple blunting and phosphorylation of DNA at once Universal - preparing any kind of DNA for blunt-end ligation Premium reagents - reproducible results PRODUCT SPECIFICATIONS Optimum activity at room temperature, around 25 C Inactivation at 75 C for 20 min Up to 1-5 microgram of the linear DNA can be blunted and phosphorylated in one 20 min reaction. After the thermal inactivation the reaction mixture can be used for blunt-end ligations. HighEnd Repair Kit is an ideal choice for preparing for ligations the PCR products obtained with high fidelity polymerases like (HLE0201). Prepare a 25 µl reaction: Linear DNA in TE buffer or water 1 µg of 1 kb linear DNA has ~3 pmol ends 1 µg of 0.5 kb linear DNA has ~6 pmol ends 1 µg of 0.1 kb linear DNA has ~30 pmol ends 10X HighEndBuffer 2.5 µl 1 mm dntp Mix 2.5 µl DNase-free water (PCR Water, WAT0110) HighEnd Repair Blend, 1 r/µl up to 1 µg (up to 30 pmol ends) to 24 µl Mix well; incubate for min at 25 C. 1 µl (max 2 µl) Inactivate enzymes at 75 C for 20 min and keep cooled in case the ligation is performed immediately or keep frozen in case the ligation is performed later. Alternatively, re-purify the DNA using PCR clean-up spin column kit, elute in 25 µl of water or TE and keep frozen. For subsequent ligation and cloning follow the recommendations for Rally Rapid Ligation Kit (RLK0101). 34 Info Tel: info@highqu.com

37 DNA Electrophoresis Ladders DNL0102 DNL appl. 200 appl. 2 x 0.5 ml - Take5 2 x 1 ml - Take5 2 x 0.5 ml - Take5 2 x 1 ml - Take5 Ready to load ladders contain highly purified PCR products combined with plasmid digests, they are supplied in 1x loading dye: 10 mm Tris- HCl (ph 8.0) 10 mm EDTA, glycerol and tracking dyes. Loading Dye has the same composition like the ladders, just it is 6X DNL appl. 2 x 0.5 ml - Take5 2 x 1 ml - Take56X more concentrated and contains all three tracking dyes. 1 µl of 6X dye shall be used for 5 µl of DNA sample, mixed well and loaded. DNL appl. 2 x 0.5 ml - Take5HR DNA Ladder 2 x 1 ml - Take5 DNA size determination and approximate DNA quantification on agarose gels Room-temperature-stable, always ready to be used Sharp bands, bright reference bands, indicated DNA mass Take5 ladders are supplied with loading dye for DNA samples Take5 Ladder Take5HR DNA Ladder 100 bp- 10 kb Range 0.5; 1.5 & 3 kb Reference 19 Bands 172 µg/µl Concentration Ready-to-use: Load: 5 µl/well Tracking Dyes: Xylene cyanol FF (~4 kb) Bromophenol blue (~0.4 kb) Orange G (<50 bp) 250 bp- 25 kb Range 1 & 3 kb Reference 14 Bands 104 µg/µl Concentration Ready-to-use: Load: 5 µl/well Tracking Dyes: Xylene cyanol FF (~4 kb) Bromophenol blue (~0.4 kb) Stable: Room temp.,6 months +4 C, 12 months -20 C, 24 months Stable: Room temp., 6 months +4 C, 12 months -20 C, 24 months Take5p DNA Ladder Take5p DNA Ladder 50 bp- 1.5 kb Range 0.2 & 0.5 kb Reference 17 Bands 112 µg/µl Concentration Ready-to-use: Load: 5 µl/well Tracking Dyes: Orange G (<50 bp) Stable: Room temp., 6 months +4 C, 12 months -20 C, 24 months 100 bp- 3 kb Range 0.5 & 1.5 kb Reference 12 Bands 108 µg/µl Concentration Ready-to-use: Load: 5 µl/well Tracking Dyes: Xylene cyanol FF (~4 kb) Bromophenol blue (~0.4 kb) Orange G (<50 bp) Stable: Room temp., 6 months +4 C, 12 months -20 C, 24 months order@highqu.com Order Tel:

38 Protein Electrophoresis Ladders CAT SIZE COMPONENTS COMPONENT COMPOSITION PRL0102 PRL appl. 200 appl. 2 x 0.5 ml - Ladder 2 x 0.5 ml - Protein Ladder Ready to load ladders contain highly purified prestained proteins, supplied denatured in 20 mm Tris-Phosphate (ph 7.5 at 25 C), 2% SDS, 0.2 mm DTT, 3.6 M Urea, 15% (v/v) glycerol). Approximate protein molecular weight determination on denaturing gels and Western blots Monitoring of electrophoresis process and transfer efficiency Room-temperature-stable, always ready to be used Sharp bands, bright colors CozyPrestained Protein Ladder kda Range 25 & 75 kda References 10 Bands µg/µl Concentration of each protein Ready-to-use: Load: 3-5 µl/ gel well for electrophoresis Load: 2-3 µl/ gel well for Western transfers Stable: Room temp., short term (2 weeks) +4 C, 3 months -20 C, 12 months The picture shows guidelines for approximate protein MW estimation. The apparent MW of each protein was determined by callibrating it against unstained protein of same size. For precise sizing, such calibration shall be done exactly at conditions used kda Range 25 & 75 kda References 13 Bands µg/µl Concentration of each protein Ready-to-use: Load: 3-5 µl/ gel well for electrophoresis Load: 2-3 µl/ gel well for Western transfers Stable: Room temp., short term (2 weeks) +4 C, 3 months -20 C, 12 months The picture shows guidelines for approximate protein MW estimation. The apparent MW of each protein was determined by callibrating it against unstained protein of same size. For precise sizing, such calibration shall be done exactly at conditions used. 36 Info Tel: info@highqu.com