contents PCR, RT-PCR & Real-time PCR RT-PCR Real-time PCR PCR Reagents Selection Chart of PCR Reagents 1 Selection Chart of RT-PCR Systems 18

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3 contents PCR, RT-PCR & Real-time PCR PCR Reagents Selection Chart of PCR Reagents 1 Taq DNA Polymerase 3 Pfu DNA Polymerase 4 Fast HiFidelity PCR Kit 5 Taq Plus DNA Polymerase 6 HotMaster Taq DNA Polymerase 7 Long Taq DNA Polymerase 8 Taq Platinic DNA Polymerase 9 Multi PCR Kit 10 2 PCR MasterMix 11 2 GC-Rich PCR MasterMix 12 2 HotStart Taq PCR MasterMix 12 2 HotMaster Taq PCR MasterMix 12 Golden Fast PCR System 13 Golden Easy PCR System PCR Reagents Individual Buffers and Accessories 17 RT-PCR Selection Chart of RT-PCR Systems 18 Quant Reverse Transcriptase 19 QuantScript RT Kit 20 Quant One Step RT-PCR Kit 20 FastQuant RT Kit (With gdnase) 21 TIANScript M-MLV 22 TIANScript RT Kit 23 FastLine Cell cdna Kit Real-time PCR Selection Chart of Real-time PCR Products 26 RealMasterMix (SYBR Green) 27 SuperReal PreMix Plus(SYBR Green) 28 RealMasterMix (Probe) 29

4 contents Quant One Step qrt-pcr (SYBR Green) Kit 30 Quant One Step qrt-pcr (Probe) Kit 31 Quant qrt-pcr (SYBR Green) Kit mirna Products mircute mirna First-strand cdna Synthesis Kit 34 mircute mirna qpcr Detection Kit(SYBR Green) 35 DNA Molecular Weight Marker DNA Molecular Weight Marker List 37 Other Products List Genomic DNA Purification Products 38 RNA Purification Products 39 Plasmid DNA Purification Products 39 PCR Cleanup & Gel Extraction Products 40 Cloning Products 41

5 PCR Product Part II PCR, RT-PCR & Real-time PCR PCR Reagents Selection Chart of PCR Reagents Standard ET101 Taq DNA Polymerase ET105 Taq Plus DNA Polymerase High Efficiency and Fidelity For Long PCR Products ET103 Long Taq DNA Polymerase Classic Reaction Set-up High Efficiency, Fidelity and Heat-stable ET104 Taq Platinic DNA Polymerase EP101 Pfu DNA Polymerase High Fidelity PCR Product KP202 Fast HiFidelity PCR Kit PCR Reagents High Specificity ET106 HotMaster Taq DNA Polymerase Multi PCR KT109 Multi PCR Kit High Sensitivity and Convenient 2 MasterMix Fast and High Specificity KT501 Golden Fast PCR Kit Convenient Set-up Golden PCR System High Cost-performance KT221 Golden Easy PCR System KG201 TIANcombi DNA Lyse&Amp PCR Kit Combination of DNA Extraction and PCR KG202 GMO Crop Extraction & Amplification PCR Kit 1

6 PCR Product PCR Product Pfu DNA Ploymerase Taq DNA Ploymerase Long Taq DNA Ploymerase Taq Platinic DNA Ploymerase Taq Plus DNA Ploymerase HotMaster Taq DNA Ploymerase 2 Pfu PCR MasterMix 2 Taq PCR MasterMix 2 LongTaq DNA Ploymerase 2 Taq Platinic PCR MasterMix 2 Taq Plus PCR MasterMix 2 GC-Rich PCR MasterMix 2 HotMaster PCR MasterMix ep101 ET101 ET103 ET104 ET105 ET106 KP201 KT201 KT203 KT204 KT205 KT206 KT208 Excellent amplification Fragment Length Possible amplification Fragment Length < 2 kb < 6 kb < 15 kb < 4 kb < 10 kb < 6 kb < 2 kb < 6 kb < 15 kb < 4 kb < 10 kb < 10 kb < 6 kb < 4 kb < 8 kb < 30 kb < 6 kb < 20 kb < 8 kb < 4 kb < 8 kb < 30 kb < 6 kb < 20 kb < 20 kb < 8 kb Efficiency Fidelity GC-rich '-Terminal A Convenience Product couldn't be used in high effective TA clone directly. Product could be used in high effective TA clone directly. Product could be used in TA clone directly but for the high effective cloning 3'-Terminal A need to be added. 2

7 PCR Product Taq DNA Polymerase Standard and Specialized PCR Applications TIANGEN Taq DNA Polymerase is purified from E.coli expressing a cloned Thermus aquaticus DNA polymerase gene. The enzyme consists of a single polypeptide with a molecular weight of approximately 94 kda. This enzyme has a 5' 3' DNA polymerase and a 5' 3' exonuclease activity but lacks a 3' 5' exonuclease activity, which results in a 3'-dA overhangs PCR product. TIANGEN Taq DNA Polymerase can amplify DNA target up to 6 kb, with the velocity of 1~2 kb/min. Taq DNA polymerase is heat-stable and will synthesize DNA at elevated temperatures from single-stranded templates in the presence of a primer. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form within 30 minutes at 74 C using hering sperm DNA as template. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases is confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN Taq DNA Polymerase Specifications Concentration Recombinant Enzyme 2.5 U/µl, 5 U/µl Extension Range < 6 kb Extension Rate 1-2 kb/min at Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases Applications Standard PCR Yes Yes Yes No no no no Product Name ET Taq DNA Polymerase 250 U, 2.5 U/µl ET (10 Taq Buffer mixed 250 U, 5 U/µl ET with Mg 2+ ) 500 U, 2.5 U/µl ET U, 5 U/µl ET U, 2.5 U/µl ET Taq DNA Polymerase 250 U, 5 U/µl ET (10 Taq Buffer and Mg 2+ ) 500 U, 2.5 U/µl ET U, 5 U/µl KT Taq PCR MasterMix 1 ml KT (with loading dye) 5 1 ml KT Taq PCR MasterMix 1 ml KT (without loading dye) 5 1 ml KT Golden Easy PCR System I : 250 U; II : 5 ml KT (with loading dye) I : 500 U; II : 10 ml 10 Taq Buffer Experimental Example Polymerase Storage Buffer 200 mm Tris-HCl (ph 8.4) 20 mm Tris-HCl (ph 8.0) 200 mm KCl 0.1 mm EDTA 15 mm MgCl 2 1 mm DTT Others 100 mm KCl Store at -20 C 50% glycerol Stabilizers Template: Human genomic DNA Target fragment: 1 kb Add the following components to a sterile PCR tube placed on ice: Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 Taq Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl Taq (2.5 U/µl) µl ddh 2 O Up to 50 µl PCR Product Perform 30 cycles of PCR amplification as follows: 94 3 min s s 30 Cycles 72 1 min 72 5 min 3

8 PCR Product Pfu DNA Polymerase High Fidelity PCR Amplification PCR Product Pfu DNA Polymerase is designed for the high-fidelity PCR amplification. High fidelity is provided by a proprietary enzyme preparation containing recombinant DNA polymerase from E.coli with proofreading activity (Pyrococcus furiosis DNA Polymerase gene). Unlike Taq DNA polymerase, highly thermostable Pfu DNA polymerase possesses 3' 5' exonuclease proofreading activity which enables the polymerase to correct nucleotide-misincorporation errors. This means that Pfu DNA polymerase-generated PCR fragments have fewer errors than the Taq-generated PCR inserts. Polymerase provides much higher fidelity than Taq DNA polymerase when doing PCR of problematic and / or GC-rich templates. Pfu DNA polymerase is superior for techniques that require highfidelity DNA synthesis. TIANGEN Pfu DNA polymerase could maintain 90% activity after 1-hour incubation at 95 C. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form within 30 minutes at 74 C using hering sperm DNA as template. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases is confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN Pfu DNA Polymerase Specifications Concentration Recombinant Enzyme Extension Range Extension Rate 5 3 Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases 2.5 U/µl Yes < 2 kb kb/min No no Yes no no no Product Name EP Pfu DNA Polymerase 250 U, 2.5 U/µl EP (10 Pfu Buffer mixed 500 U, 2.5 U/µl with Mg 2+ ) KP Pfu PCR MasterMix 0.5 ml KP (with loading dye) 5 1 ml KP Pfu PCR MasterMix 0.5 ml KP (without loading dye) 5 1 ml 10 Taq Buffer Experimental Example Polymerase Storage Buffer 200 mm Tris-HCl (ph 8.8) 50 mm Tris-HCl (ph 8.2) 100 mm KCl 0.1 mm EDTA 100 mm (NH 4 ) 2 SO 4 1 mm DTT 20 mm MgSO 4 50% glycerol Others Stabilizers Store at -20 C. Template: Genomic DNA Target fragment: 1 kb Add the following components to a sterile PCR tube sitting on ice: Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 Pfu Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl Pfu (2.5 U/µl) µl ddh 2 O Up to 50 µl Perform 30 cycles of PCR amplification as follows: 94 3 min s s 30 Cycles 72 2 min 72 5 min 4

9 PCR Product Fast HiFidelity PCR Kit For fast, highly sensitive and reliable high-fidelity PCR Fast HiFidelity DNA Polymerase is purified from the hyper thermophilic original strain Pyrococcus spp, with super 3 '- 5' exonuclease activity (Proofreading activity). This product uses innovative synthetic affinity ligands technology, ligand inhibits polymerases and exonuclease activity in a temperature dependent manner; ligand A inhibits the DNA Polymerase activity at room temperature, which effectively reducing nonspecific amplification; ligand B inhibits the 3' 5 exonuclease activity and prevents the primer and template from degrading, providing highly sensitive and reliable high-fidelity PCR. The kit also contains PCR Enhancer, a unique additive enabling efficient amplification of difficult templates. KP μl 50 rxn KP μl 200 rxn KP μl 1000 rxn Components 50 μl 50 rxn 50 μl 200 rxn 50 μl 1000 rxn Fast HiFidelity Polymerase 125 U 500 U 5X500 U 5 Fast HiFidelity PCR Buffer* 500 µl 2 1 ml 10 1 ml 20 Fast PCR Enhancer 500 µl 500 µl µl 5 DNA Loading Buffer 500 µl 1 ml 5 1 ml *contains dntps, ligand a/b, mg 2+, and stabilizers Store at -20 Features High Fidelity: Owing to its high proofreading activity, fidelity of Fast HiFidelity Polymerase is more than 80 times than that of Taq DNA polymerase, 8 times of Pfu DNA Polymerase. High Amplifying Speed: Fast HiFidelity Polymerase has a unique extension enhancement factor, which make the extending speed and the amplification significantly improved, extension speed can be increased to 15~30 sec/kb, 2~4 times faster than Taq DNA Polymerase, 4~8 times faster than Pfu DNA Polymerase. Easy to Operation: This product uses innovative synthetic affinity ligands technology, making it easy to perform the highly specific hot Start PCR without any extra steps. Mg 2+ Automatic Balanced System: Owing to its uniquely balanced system for Mg 2+ concentration, optimization of Mg 2+ concentrations in PCR often minimal or not required. Experimental Example m: tiangen marker iii 1: mpkc (2.0 kb) 2: Hn2.0 (2.0 kb) 3: Hn4.5 (4.5 kb) the result show that tiangen Taq plus and t company of other brand is difficult to effectively amplify the above samples at the speed of sec/kb, but Fast HiFidelity PcR Kit could reach 15 sec/kb, and the yield will be increased at the speed of 30 sec/kb, which is recommended. PCR Product 5

10 PCR Product PCR Product Taq Plus DNA Polymerase For High Efficiency and High Fidelity PCR Reaction Taq Plus DNA Polymerase is provided as a mixture of Taq DNA Polymerase and the proofreading enzyme Pfu DNA Polymerase. The product possesses both the processivity of Taq and the high fidelity of Pfu. The specially formulated Taq Plus Polymerase allows amplification of the higher fidelity and longer templates than the single-enzyme formulation. It is a better choice for amplifying complex template, such as GC-rich template. It has a 5 3 Exonuclease activity as well as 3 5 Exonuclease activity. Taq Plus DNA Polymerase is designed for enabling reliable amplification of DNA target up to 10 kb. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form in 30 minutes at 74 C using hering sperm DNA as substrate. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN Taq Plus DNA Polymerase Specifications Concentration Mixture Enzyme Extension Range 5 3 Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases Applications Amplification of complicated sencondary structure. 2.5 U/µl Yes < 10 kb Yes Yes Yes no no no Product Name ET Taq Plus DNA Polymerase 250 U, 2.5 U/µl (10 Taq Plus Buffer ET mixed with Mg 2+ ) 500 U, 2.5 U/µl KT Taq Plus PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Taq Plus PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml 10 Taq Plus Buffer Experimental Example Polymerase Storage Buffer 200 mm Tris-HCl (ph 8.4) 20 mm Tris-HCl (ph 8.0) 200 mm KCl 0.1 mm EDTA 100 mm (NH4) 2 SO 4 1 mm DTT 15 mm MgCl mm KCl Others 50% glycerol Store at -20 C. Template: Genomic DNA Target fragment: 1 kb Stabilizers Add the following components to a sterile PCR tube sitting on ice: Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 Taq Plus Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl Taq Plus (2.5 U/µl) µl ddh 2 O Up to 50 µl Perform 30 cycles of PCR amplification as follows: 94 3 min s s 30 Cycles 72 1 min 72 5 min 6

11 PCR Product HotMaster Taq DNA Polymerase Innovative and Unique Hot-Start / Cold-Stop Taq DNA Polymerase HotMaster Taq DNA Polymerase is a superior alternative for performing PCR experiments generally known as hot start PCR. HotMaster blocks the substrate binding sites of DNA polymerases in a temperature-dependent manner. Inactive polymerase-inhibitor complexes are formed at temperatures < 40 C, when the affinity of HotMaster for Taq polymerase is higher than the binding affinity to with of the template DNA. Between 40 C and 55 C, the HotMaster competes with the template DNA for binding to the Taq polymerase, thereby shifting the binding equilibrium towards complex formation with only target-specific primed template DNA. At temperatures above 55 C the HotMaster inhibitor is displaced from complexes with the Taq polymerase by target-specific primed template DNA. A unique performance feature of the HotMaster inhibitor is that it can go through multiple temperature cycles of binding-equilibrium competition-dissociation during PCR without irreversible heat inactivation. Where other Taq polymerase formulations for hot start PCR block the activity of Taq polymerase only prior to the first high temperature step, the TIANGEN HotMaster provides sustained temperature control throughout PCR. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntp into an acid-insoluble form in 30 minutes at 65 C using hering sperm DNA as substrate. Product Name HotMaster Taq DNA ET U, 2.5 U/µl Polymerase ET (10 HotMaster Taq Buffer 500 U, 2.5 U/µl mixed with Mg 2+ ) KT HotMaster PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT HotMaster PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml 10 HotMaster Taq Buffer 200 mm Tris-HCl (ph 8.4) 200 mm KCl 100 mm (NH4) 2 SO 4 25 mm MgCl 2 Others Polymerase Storage Buffer 20 mm Tris-HCl (ph 8.0) 0.1 mm EDTA 1 mm DTT 100 mm KCl 50% glycerol Stabilizers Store at -20 C. PCR Product Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN HotMaster Taq DNA Polymerase Specifications Concentration Recombinant Enzyme Extension Range 5 3 Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases Applications Low copy detection PCR. DNA sequencing. Multiplex PCR. TA clone. 2.5 U/µl Yes < 6 kb Yes Yes No no no no Experimental Example Template: Genomic DNA Target fragment: 1 kb Add the following components to a sterile PCR tube sitting on ice: Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 HotMaster Taq Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl HotMaster Taq (2.5 U/µl) µl ddh 2 O Up to 50 µl Perform 30 cycles of PCR amplification as follows: 94 3 min s s 30 Cycles 65 1 min 65 5 min 7

12 PCR Product Long Taq DNA Polymerase Effective and Accurate Long-Length PCR Product Name Long Taq DNA ET U, 2.5 U/µl Polymerase PCR Product TIANGEN Long Taq DNA Polymerase is a mix of Taq DNA polymerase and a powerful high-fidelity enzyme to ensure high amplification efficiency and maximum fidelity. Long Taq DNA Polymerase has a 3 5 Exonuclease Activity. Two different unique PCR buffers are suitable for different templates and enhance extension rates and fidelity, even when amplifying complex templates. PCR products of up to 40 kb can be amplified reliably. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form in 30 minutes at 72 C using hering sperm DNA as substrate. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN Long Taq DNA Polymerase Specifications Concentration Mixture Enzyme Extension Range 5 3 Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases 2.5 U/µl Yes < 40 kb Yes no Yes no no no (10 Long Taq Buffer ET mixed with Mg 2+ ) KT Long Taq PCR 0.5 ml MasterMix KT (with loading dye) 5 1 ml KT Long Taq PCR 0.5 ml MasterMix KT (without loading dye) 5 1 ml 10 Long Taq Buffer We provide two different buffers: 10 Long Taq Buffer I 10 Long Taq Buffer II Polymerase Storage Buffer 20 mm Tris-HCl (ph 8.0) 0.1 mm EDTA 1 mm DTT 100 mm KCl 50% glycerol Stabilizers Store at -20 C. Experimental Example Template: Genomic DNA Target fragment: 1 kb Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 Long Taq Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl Long Taq (2.5 U/µl) µl ddh 2 O Up to 50 µl U, 2.5 U/µl Add the following components to a sterile PCR tube sitting on ice: Perform 30 cycles of PCR amplification as follows: 94 3 min s s 30 Cycles 72 1 min 72 5 min 8

13 PCR Product Taq Platinic DNA Polymerase Highly Reliable End-Point PCR Applications with Unrivalled Ease-of-Use Taq Platinic DNA Polymerase is ideal for automatic hot-start amplification of DNA fragments with improved specificity over Taq DNA polymerase. It is derived from recombinant Taq DNA polymerase by binding of a thermolabile inhibitor containing chemical compound to Taq DNA polymerase. Taq Platinic DNA Polymerase has a 3 5 Exonuclease Activity as well as 5 3 Exonuclease Activity. During the initial denaturation step of PCR, the inhibitor is denatured and active Taq DNA polymerase is released into the reaction. With Taq Platinic DNA Polymerase, improved specificity and high yields of PCR compared to Taq DNA polymerase without hot-start could be reached. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form in 30 minutes at 74 C using hering sperm DNA as substrate. Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests. Functionally tested in PCR. TIANGEN Taq Platinic DNA Polymerase Specifications Concentration Mixture Enzyme Extension Range 5 3 Exonuclease Activity Extra "A" Addition 3 5 Exonuclease Activity Contaminating Nucleases Contaminating RNases Contaminating Proteases 2.5 U/µl Yes < 4 kb Yes Yes Yes no no no Product Name Taq Platinic DNA ET Polymerase 250 U, 2.5 U/µl (10 Taq Platinic Buffer ET mixed with Mg 2+ ) 500 U, 2.5 U/µl KT Taq Platinic PCR 0.5 ml MasterMix KT (with loading dye) 5 1 ml KT Taq Platinic PCR 0.5 ml MasterMix KT (without loading dye) 5 1 ml 10 Taq Platinum Buffer Buffer I Buffer II 200 mm Tris-HCl (ph 8.4) 200 mm Tris-HCl (ph 8.8) 200 mm KCl 100 mm KCl 100 mm (NH 4 ) 2 SO mm (NH 4 ) 2 SO 4 15 mm MgCl 2 20 mm MgCl 2 Others others Polymerase Storage Buffer 20 mm Tris-HCl (ph 8.0) 0.1 mm EDTA 1 mm DTT 100 mm KCl 50% glycerol Stabilizers Store at -20 C. Experimental Example Template: Genomic DNA Target fragment: 1 kb Add the following components to a sterile PCR tube sitting on ice: Template < 1 µg Primer 1 (10 µm) 1 µl Primer 2 (10 µm) 1 µl 10 Taq Platinic Buffer 5 µl dntp Mixture (2.5 mm each) 4 µl Taq Platinic (2.5 U/µl) µl ddh 2 O Up to 50 µl PCR Product Perform 30 cycles of PCR amplification as follows: 94 5 min s s 30 Cycles 72 2 min 72 5 min 9

14 PCR Product Multi PCR Kit For highly specific and sensitive multiplex PCR without optimization PCR Product TIANGEN Multi PCR Kit is specifically developed for multiplex PCR. TIANGEN Multi PCR Kit provides Multi HotStart DNA Polymerase with a unique Multi HotStart PCR buffer containing the novel synthetic Factor MP. Together with optimized salt concentrations, this Factor stabilizes specifically template bound primers and enables efficient extension of all primers in the reaction without optimization. This chemically modified HotStart Taq DNA polymerase is inactive during reaction setup and only becomes active after incubation at 95ºC, preventing the formation of nonspecific products and primer dimers in initial and later PCR cycles. Multiplex assays can be set up at room temperature and HotStart Taq DNA Polymerase in the master mix is then activated by a 15-minute incubation at 95 C, which can be incorporated into any existing thermal-cycler program. Features Chemically Modified Polymerase: Only becomes active after incubation at 95ºC for 15 min High Performance: High specificity and sensitivity with a built-in hot start DNA Polymerase High Sensitivity: Highly suited for many types of multiplex PCR applications Convenience: Easy to use and cost-effective Applications KT rxn (250 U) KT rxn (500 U) Components 50 rxn 100 rxn Multi HotStart Polymerase 250 U 500 U 10 Multi HotStart PCR Buffer 1.8 ml 1.8 ml Super pure dntp (2.5mM each) 240 µl 500 µl MgCl 2 (mm) 1.8 ml 1.8 ml ddh 2 O 2 1 ml 5 ml Store at -20 C. Experimental Example Template: Genomic DNA Target fragment: 7 fragments(100 bp bp) Add the following components to a sterile PCR tube sitting on ice: Templates 100 ng 10 primer mix (2 µm each) 5 µl 10 Multi HotStart Buffer 5 µl dntps 200 µm Multi HotStart Polymerase (5 U/µl) 1 µl ddh 2 O Up to 50 µl Amplification of Multiplex PCR regions. Amplification of complicated templates. Amplification and analysis with high specificity. Definition of Activity Unit One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dntps into an acid-insoluble form in 30 minutes at 74 C using hering sperm DNA as substrate. Perform 40 cycles of PCR amplification as follows: min s s 40 Cycles s min Notice: Duration time when using Multi PCR Kit should be based on the target fragments: 60 s for fragments smaller than 500 bp 90 s for fragments from 500 bp to 1500 bp 120 s for fragments longer than 2000 bp This Chemically Modified Polymerase only becomes active after incubation at 95ºC for 15 min 10

15 PCR Product 2 PCR MasterMix Premixed Solution for Convenient PCR Setup TIANGEN PCR MasterMix is a kind of ready-to-use mixture of DNA polymerase, salts, magnesium, and dntps for efficient PCR amplification. All you have to do is adding template and primers, reducing set-up time by half. TIANGEN PCR MasterMixes provide a variety of enzyme formulations as well as reagents licensed and qualified for PCR. Features Higher Sensitivity: High PCR specificity and reduced nonspecific amplification Convenience: Minimal optimization and easy to setup reaction Quality Control The absence of endodeoxyribonucleases, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests, functionally tested in PCR. Content and Storage Each PCR MasterMix is supplied as 2 concentration and contains DNA polymerase salts, magnesium and dntps. PCR MasterMix could be stored at 4 for 3 months and should be stored at -20 for more than one year. Product Name KT Taq PCR MasterMix 1 ml KT (with loading dye) 5 1 ml KT Taq PCR MasterMix 1 ml KT (without loading dye) 5 1 ml KP Pfu PCR MasterMix 0.5 ml KP (with loading dye) 5 1 ml KP Pfu PCR MasterMix 0.5 ml KP (without loading dye) 5 1 ml KT Taq Plus PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Taq Plus PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml KT Hotstart Taq PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Hotstart Taq PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml KT Long Taq PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Long Taq PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml KT Taq Platinic PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Taq Platinic PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml KT GC-Rich PCR MasterMix (with loading dye) 0.5 ml KT206 2 GC-Rich PCR MasterMix (without loading dye) 0.5 ml KT HotMaster PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT HotMaster PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml PCR Product Experimental Example Add the following components to a sterile PCR tube: Template < 1 µg Primer 1 (10 µm) 2 µl Primer 2 (10 µm) 2 µl 2 MasterMix 25 µl ddh 2 O Up to 50 µl The final concentration of components in TIANGEN MasterMix: 10 mm Tris-HCl (ph 8.3) 50 mm KCl 1.5 mm MgCl µm dntp each 0.05 U / µl Polymerase ddh 2 O Other Stabilizers and Enhancers 11

16 PCR Product 2 GC-Rich PCR MasterMix Premixed Solution Especially for GC-Rich Templates PCR 2 GC-Rich PCR MasterMixes is supplied in 2 concentration and contains Taq Plus DNA polymerase, salts, magnesium, dntps, PCR enhancers and buffers. With 2 GC-Rich PCR MasterMix set-up time could be reduced by half. PCR products of up to 20 kb can be reliably amplified (up to 10 kb for complex templates). Product Name KT GC-Rich PCR MasterMix 0.5 ml (with loading dye) KT GC-Rich PCR MasterMix 0.5 ml (without loading dye) Store at -20 C. Features PCR Product High efficiency: Blend of DNA polymerases allows automatic hot-start High Fidelity: especially for high-specificity, high-fidelity DNA amplification applications Convenience: Ready-to-use master mix needs minimal pipetting steps 2 HotStart Taq PCR MasterMix Premixed Solution Using HotStart Taq Polymerase for High PCR Specificity HotStart Taq PCR MasterMix contains HotStart Taq DNA Polymerase, the unique PCR Buffer that minimizes the requirement for optimization, as well as dntps. Providing all components in a master mix reduces pipetting steps and risk of contamination, while increasing throughput and reproducibility. Product Name KT Hotstart Taq PCR MasterMix 0.5 ml KT (with loading dye) 5 1 ml KT Hotstart Taq PCR MasterMix 0.5 ml KT (without loading dye) 5 1 ml 12

17 PCR Product Golden Fast PCR System Fast, Speciffic and Efficiency two-component PCR System Dual-component simple PCR system contains Golden Fast DNA Polymerase and 2 Golden Fast Reaction Mix. Golden Fast DNA Polymerase is heat-resistant DNA polymerase designed for fast PCR. It can extend 1Kb in 15sec which is time-saving and anneal within 5s. The system is fast, highly specific, sensitive and stable which could reduce human factor error. It could be applied in regular PCR, secondary template, and high throughput gene detection. Experiment Procedure Product Name KT Golden Fast PCR Kit Golden Fast PCR Kit KT (without loading dye) 1 ml 5 ml Components KT KT Golden Fast Reaction Mix 1 ml Golden Fast DNA polymerase 100 U 5 ml 500 U Store at -20 C. PCR Product Experimental Example Fragment Length 5kb 2kb 1kb 500bp 0 50 min 100 min 150 min 200 min Time Golden Fast PCR Kit Taq DNA Polymerase Amplification time comparison between Fast PCR and traditional PCR 13

18 PCR Product Golden Easy PCR System PCR System with two components Features Product Name KT Golden Easy PCR System I: 250 U; II: 5 ml KT (with loading dye) I: 500 U; II: 10 ml PCR Product Stability: The system contains all needed components; Unique reaction mixes make it stable Convenience: All you have to do is to mix two components and add template and primers. Applications Golden Easy PCR Systems provide high sensitivity and variety of applications Loading Dye: PCR products can be directly used for electrophoresis after amplification. Experiment Procedure Step 1: Mix two components, template and primers; Step 2: Start the reaction; Step 3: Perform the electrophoresis after PCR; Step 4: Perfect result. Component I Component II Template & Primers Components KT KT Golden DNA Polymerase (I) 250 U 500 U 2 Reaction Mix (II) 5 ml 10 ml Store at -20 C. Experimental Example Template: Genomic DNA Target fragment: 1 kb Add the following components to a sterile PCR tube sitting on ice: Template < 1 µg Primer 1 (10 µm) 2 µl Primer 2 (10 µm) 2 µl Golden DNA polymerase(2.5 U/µl) 0.5 µl 2 Reaction Mix 25 µl ddh 2 O Up to 50 µl 14

19 PCR Product Product Name EP EP EP EP ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET ET Pfu DNA Polymerase 250 U, 2.5 U/µl (10 Pfu Buffer mixed with Mg 2+ ) Pfu DNA Polymerase 500 U, 2.5 U/µl (10 Pfu Buffer mixed with Mg 2+ ) Pfu DNA Polymerase 250 U, 2.5 U/µl (10 Pfu Buffer and Mg 2+ ) Pfu DNA Polymerase 500 U, 2.5 U/µl (10 Pfu Buffer and Mg 2+ ) Taq DNA Polymerase 250 U, 2.5 U/µl (10 Taq Buffer mixed with Mg 2+ ) Taq DNA Polymerase 250 U, 5 U/µl (10 Taq Buffer mixed with Mg 2+ ) Taq DNA Polymerase 500 U, 2.5 U/µl (10 Taq Buffer mixed with Mg 2+ ) Taq DNA Polymerase 500 U, 5 U/µl (10 Taq Buffer mixed with Mg 2+ ) Taq DNA Polymerase 250 U, 2.5 U/µl (10 Taq Buffer and Mg 2+ ) Taq DNA Polymerase 250 U, 5 U/µl (10 Taq Buffer and Mg 2+ ) Taq DNA Polymerase 500 U, 2.5 U/µl (10 Taq Buffer and Mg 2+ ) Taq DNA Polymerase 500 U, 5 U/µl (10 Taq Buffer and Mg 2+ ) Long Taq DNA Polymerase 250 U, 2.5 U/µl (10 Long Taq Buffer mixed with Mg 2+ ) Long Taq DNA Polymerase 500 U, 2.5 U/µl (10 Long Taq Buffer mixed with Mg 2+ ) Long Taq DNA Polymerase 250 U, 2.5 U/µl (10 Long Taq Buffer and Mg 2+ ) Long Taq DNA Polymerase 500 U, 2.5 U/µl (10 Long Taq Buffer and Mg 2+ ) Taq Platinic DNA Polymerase 250 U, 2.5 U/µl (10 Taq Platinic Buffer mixed with Mg 2+ ) Taq Platinic DNA Polymerase 500 U, 2.5 U/µl (10 Taq Platinic Buffer mixed with Mg 2+ ) Taq Platinic DNA Polymerase 250 U, 2.5 U/µl (10 Taq Platinic Buffer and Mg 2+ ) Taq Platinic DNA Polymerase 500 U, 2.5 U/µl (10 Taq Platinic Buffer and Mg 2+ ) Taq Plus DNA Polymerase 250 U, 2.5 U/µl (10 Taq Plus Buffer mixed with Mg 2+ ) Taq Plus DNA Polymerase 500 U, 2.5 U/µl (10 Taq Plus Buffer mixed with Mg 2+ ) Taq Plus DNA Polymerase 250 U, 2.5 U/µl (10 Taq Plus Buffer and Mg 2+ ) Taq Plus DNA Polymerase 500 U, 2.5 U/µl (10 Taq Plus Buffer and Mg 2+ ) PCR Product 15

20 PCR Product PCR Product Product Name ET HotMaster Taq DNA Polymerase (10 HotMaster Taq Buffer mixed with Mg 2+ ) 250 U, 2.5 U/µl ET HotMaster Taq DNA Polymerase (10 HotMaster Taq Buffer mixed with Mg 2+ ) 500 U, 2.5 U/µl KP Pfu PCR MasterMix (with loading dye) 0.5 ml KP Pfu PCR MasterMix (with loading dye) 5 1 ml KP Pfu PCR MasterMix 0.5 ml KP Pfu PCR MasterMix 5 1 ml KP Fast HiFidelity PCR Kit 50 µl 50rxn KP Fast HiFidelity PCR Kit 50 µl 200rxn KP Fast HiFidelity PCR Kit 50 µl 1000rxn KT Multi PCR Kit (250 U) KT Multi PCR Kit 100 preps (500 U) KT Taq PCR MasterMix (with loading dye) 1 ml KT Taq PCR MasterMix (with loading dye) 5 1 ml KT Taq PCR MasterMix 1 ml KT Taq PCR MasterMix 5 1 ml KT Hotstart Taq PCR MasterMix (with loading dye) 0.5 ml KT Hotstart Taq PCR MasterMix (with loading dye) 5 1 ml KT Hotstart Taq PCR MasterMix 0.5 ml KT Hotstart Taq PCR MasterMix 5 1 ml KT Long Taq PCR MasterMix (with loading dye) 0.5 ml KT Long Taq PCR MasterMix (with loading dye) 5 1 ml KT Long Taq PCR MasterMix 0.5 ml KT Long Taq PCR MasterMix 5 1 ml KT Taq Platinic PCR MasterMix (with loading dye) 0.5 ml KT Taq Platinic PCR MasterMix (with loading dye) 5 1 ml KT Taq Platinic PCR MasterMix 0.5 ml KT Taq Platinic PCR MasterMix 5 1 ml KT Taq Plus PCR MasterMix (with loading dye) 0.5 ml KT Taq Plus PCR MasterMix (with loading dye) 5 1 ml KT Taq Plus PCR MasterMix 0.5 ml KT Taq Plus PCR MasterMix 5 1 ml KT GC-Rich PCR MasterMix (with loading dye) 0.5 ml KT GC-Rich PCR MasterMix 0.5 ml KT Golden Easy PCR System (with loading dye) i:250 U; II:5 ml KT Golden Easy PCR System (with loading dye) i:500 U; II:10 ml KT Golden Fast PCR Kit 1 ml KT Golden Fast PCR Kit (without loading dye) 5 ml 16

21 PCR Product PCR Reagents Individual Buffers and Accessories Product Name CD102 T7 Primer 200 µl (10 µm) CD103 T3 Primer 200 µl (10 µm) CD104 SP6 Primer 200 µl (10 µm) CD105 Random Octamers 200 µl (10 µm) CD106 oligo (dt) µl (10 µm) CD Super pure dntp (2.5mM each) 0.5 ml CD Super pure dntp (2.5mM each) 1 ml CD Super pure dntp (2.5mM each) 5 1 ml CD Super pure dntp (10mM each) 0.5 ml CD Super pure dntp (10mM each) 1 ml CD Super pure dntp (10mM each) 5 1 ml CD Super pure datp (100mM each) 0.5 ml CD Super pure dgtp (100mM each) 0.5 ml CD Super pure dctp (100mM each) 0.5 ml CD Super pure dttp (100mM each) 0.5 ml CD dntp (2.5mM each) 1 ml CD dntp (2.5mM each) 5 1 ml CD dntp (10mM each) 1 ml CD dntp (10mM each) 5 1 ml RP202 PCR Enhancer 500 µl RT ddh 2 O 100 ml RT ddh 2 O 500 ml RT ddh 2 O 5 1 ml RT DNase/RNase-Free ddh 2 O 5 5 ml RT DNase/RNase-Free ddh 2 O 100 ml RT101 agarose (low EEO) 50 g RT203 ethidium Bromide Buffer ( EB BuFFER) 1 ml RT x TAE 50 ml RT x TBE 500 ml RT x DNA Loading Buffer ( Bromophenol Blue ) 5 ml RT x DNA Loading Buffer ( Bromophenol Blue, Xylene cyanol ) 5 ml PCR Product 17

22 RT-PCR Product RT*-PCR Selection Chart of RT*-PCR Systems ER103 Quant Reverse Transcriptase Reverse Transcription KR103 Quantscript RT* Kit KR106 FastQuant RT* Kit RT-PCR Product RT-PCR Products Unique Reverse Transcription RT*-PCR Cell to cdna KR113 Quant One Step RT*-PCR Kit KR105 FastLine Cell cdna Kit ER103 TIANScript M-MLV Classical Reverse Transcription KR104 TIANScript RT* Kit * RT: Reverse Transcription 18

23 RT-PCR Product Quant Reverse Transcriptase Reverse Transcription of 50 ng to 2 µg RNA per Reaction at One Temperature Quant Reverse Transcriptase is a new, unique enzyme, which is different from the reverse transcriptase of M-MLV or AMV. Quant Reverse Transcriptase is a recombinant heterodimeric enzyme expressed in E.coli. Quant Reverse Transcriptase is specially designed for all reverse transcription with any amount of RNA from 50 ng to 2 µg per reaction. Features High Performance: Sensitive detection of as few as 50 ng of template. High Yields: High cdna yields due to high-affinity enzyme. Convenience: Fast and easy procedure with no tedious pipetting steps. No RNaseH Activity: No additional RNaseH digestion step required. Accomplish the whole reaction of reverse transcription at 37. ER rxn ER rxn ER rxn Components 25 rxn 50 rxn 100 rxn Quant Reverse Transcriptase 25 µl 50 µl 2 50 µl 10 RT Buffer 70 µl 150 µl µl Store at -20 C. Experimental Example Applications RT-PCR Real-time RT-PCR Synthesis of double-stranded cdna for cloning SAGE (Serial Analysis of Gene Expression) RACE (Rapid Amplification of cdna Ends) Analysis of transcription start site by primer extension Comparison of Various Reverse Transcriptases: Reverse transcription was carried out with different reverse transcriptases according to suppliers' specifications. RT-PCR Product 19

24 RT-PCR Product Quantscript RT Kit Convenient and High Efficient Reverse Transcription of 50 ng to 2 µg RNA per Reaction Quantscript RT Kit is designed for two-step RT-PCR. The Kit contains a new and unique enzyme, Quant Reverse Transcriptase. Quant Reverse Transcriptase is specially designed for all reverse transcription with any amount of RNA from 50 ng to 2 µg per reaction. Quant Reverse Transcriptase is provided ready to use with dntps and with an optimized reaction buffer which, together with the high affinity of Quant Reverse Transcriptase for RNA, enables read-through of templates with high GC content or complex secondary structures. Features KR KR rxn 100 rxn Kit Components 25 rxn 100 rxn Quant Reverse Transcriptase 25 µl 2 50 µl Oligo(dT) 15 (10 µm) 60 µl 240 µl Random Octamers (10 µm) 60 µl 240 µl 10 RT Mix 50 µl 200 µl RNase-free ddh 2 O 1 ml 2 1 ml dntp (2.5 mm each) 60 µl 240 µl Store at -20 C. RT-PCR Product Ready to Use: The kit includes dntps, primers, buffer mix and RNase-free ddh 2 O. Unique Enzyme: The kit contains a new and unique enzyme, Quant Reverse Transcriptase. High Performance: The kit enables read-through of templates with high GC content or complex secondary structures. Applications RT-PCR Real-time RT-PCR 3 - and 5 - RACE Quant One Step RT-PCR Kit Highly Sensitive and Successful One-Step RT-PCR KR rxn The Quant One-Step RT-PCR Kit provides Quant Reverse Transcriptases, HotMaster Taq DNA Polymerase, 10 One Step RT-PCR Buffer, dntp, RNasin, and 5 RT-PCR enhancer, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates. The easily optimized components result in highly sensitive and successful results. Features One-Tube Setup: Fast and easy one-tube setup One-step RT-PCR of any RNA template without optimization High Quality: Unique enzyme mix for high specificity and sensitivity Unique Buffer: Optimized reverse-transcription and amplification buffer Kit Components 50 rxn HotMaster Taq Polymerase (2.5 U/µl) 130 µl Quant Reverse Transcriptase (for one step) 30 µl 10 RT-PCR Buffer 300 µl 5 RT-PCR enhancer 600 µl RNasin (40 U/µl) 30 µl dntp (10 mm each) 120 µl RNase-Free ddh 2 O 2 1 ml Store at -20 C. Applications RT-PCR for 10 ng to 1 µg RNA Virus RNA detection 3 - and 5 - RACE 20

25 RT-PCR Product FastQuant RT Kit (With gdnase) 21 min high efficient reverse transcription with gdna cleaning up FastQuant RT kit is an efficient, rapid and genomic DNA cleaning up reverse transcription system. This product contains gdnase which can remove genomic DNA by incubating at 42 for 3 min to protect the total RNA from genomic DNA interference. The FastQuant RT Enzyme provides a high efficient reverse transcription with 42, 15 min, and enables read-through of templates with high GC content or complex secondary structures. Due to this pre-optimization, tedious pipetting and pre-incubation steps are unnecessary, and no additional RNase H digestion step is needed. KR KR rxn 100 rxn Components 25 rxn 100 rxn 5 gdna Buffer FQ-RT Primer Mix RT Enzyme Mix 10 Fast RT Buffer RNase-free ddh2o 50 μl 50 μl 25 μl 50 μl 1 ml 200 μl 200 μl 100 μl 200 µl 2 1 ml Store at -20 C. High Reverse Transcription Efficiency: Reverse transcription efficiency above 90%. Ready to Use: Reaction system is easy to prepare, providing you to finish the whole reaction within 21min. High Performance: This product enables read-through of templates with high GC content or complex secondary structures. Good Compatibility: follow-up with real-time PCR detection, showing high sensitivity and good stability. Experimental Example Real-time analysis kit: TIAGEN SurperReal PreMix Plus (FP205) Reaction system: 20μl Applications RT-PCR Real-time RT-PCR Strong reading ability of super complex template Procedure FastQuant Rt Kit VS 21 min Fast Quant kit traditional Rt P S Other Kits 140 min Rna gdnase(3 min) gdna clean up no need of Rna purification Dnase i (30 min) Rna purification(30-40 min) Fast quant kit has higher reverse transcription ability no matter with templates contain high GC percentage or complex second structure. gdna-free Rna Quant (18 min) inactivation (3 min) Rt mmlv (60 min) inactivation (10 min) gdna-free cdna 21 RT-PCR Product Features

26 RT-PCR Product TIANScript M-MLV Reverse Transcription of 500 ng to 5 µg RNA per Reaction TIANScript M-MLV Reverse Transcriptase is an RNA-dependent DNA polymerase that can be used in cdna first strand synthesis. The enzyme is a product of the pol gene of M-MLV and consists of a single subunit with a molecular weight of 71kDa. 200 units of enzyme are used to produce cdna from 1 μg of a 1.2 kb control RNA. ER ER U (25 rxn) U (100 rxn) Kit Components 25 rxn 100 rxn TIANScript M-MLV 5 First-Strand Buffer 25 µl 150 µl 100 µl 500 µl Store at -20 C. Features Weakening activity of RNaseH : Protecting the RNA template in the maximum range. High reverse transcription activity : Increasing cdna yield and suitable for long RNA template. Stable function of enzyme Experimental Example M RT-PCR Product Applications First standard of cdna synthesis One-step RT-PCR 3 and 5 RACE Construction of cdna Library PCR amplification of 280bp and 300bp using 1µ l cdna reverse transcript with TIANGEN ER104 (lane 2, 3, 5, 6)and M-MLV from other supplier (lane 1,4) as samples. 5 µl of 20 µl PCR product per lane. M: TIANGEN MarkerⅠ. Gen Selected PCR amplification of long fragment using 1µl cdna reverse transcript with TIANGEN ER104 (lane 2, 3)and M-MLV from other supplier (lane 1) as samples. 5 ul of 20 µl PCR product per lane. M: TIANGEN MarkerⅢ. 22

27 RT-PCR Product TIANScript RT Kit Reverse Transcription of 500 ng to 5 µg RNA per Reaction TIANScript RT Kit is designed for two-step RT-PCR. The Kit contains TIANScript M-MLV Reverse Transcriptase, 5 First-Strand Buffer, a dntp mix and RNasin. Applications Frist standard of cdna synthesis Real-time qrt-pcr 3 and 5 RACE KR KR rxn 100 rxn Kit Components 25 rxn 100 rxn TIANScript M-MLV (200 U/µl) 25 µl 100 µl Oligo(dT) 15 (10 µm) 60 µl 240 µl Random (10 µm) 60 µl 240 µl 5 First-Strand Buffer 150 µl 500 µl RNase-free ddh 2 O 1 ml 2 1 ml dntp (2.5 mm each) 60 µl 240 µl RNasin (40 U/µl) 15 µl 2 30 µl Store at -20 C. RT-PCR Product 23

28 RT-PCR Product FastLine Cell cdna Kit Fast Purification of cdna from cells for PCR or Real-time PCR The kit provides a fast and simple procedure for preparing first-strand cdna directly from cultured cells without RNA purification. The kit is supplied with wash and lysis buffers for preparing lysates and stabilizing RNA, with gdna Wipeout Buffer for eliminating genomic DNA contamination, and with all reaction components for fast and efficient cdna synthesis. Features cdna synthesized by the FastLine Cell cdna Kit gives highly sensitive and reproducible results in real-time two-step RT-PCR. The kit is ideal for experiments which require a snapshot of individual transcript levels, such as: Validation of sirna-mediated gene knockdown Evaluation of drug effects Detection of gene regulation. KR rxn Kit Components 50 rxn Buffer FCW Buffer FCP gdna Wipeout Buffer (7 ) F-Quant Reverse Transcriptase Quantiscript RT Buffer (5 ) RT Primer Mix RNase-Free Water 25 ml 10 ml 100 μl 50 μl 200 μl 50 μl 1.0 ml Buffer FCW and Buffer FCP should be stored at room temperature (15-25 C); Other Reagents at -20 C. FastLine Cell cdna Kit cultured cell washing Resoluting 3~5 min 5~10 min 50 min Traditional Way Ct average Value RT-PCR Product Experimental Example 220 min cultured cell 3~5 min 60~90 min rat GAPDHI washing rat β-actin FastLine cell Kit rat GAPDH2 Rnaprep pure Kit qrt PCR result compare with the cdna produced by using FastLine Cell cdna Kit and traditional way. Sample: Rat A7r5 anchorage-dependent cell (5 x 104). Target Gene: rat GAPDH1, rat β-actin, rat GAPDH2 Result: cdna produced by FastLine Cell cdna Kit has a better pureness and sensitivity than that produced by traditional way. Rna Purification Rna gdna cleaning up 5 min gdna-free Rna Rt by Quant 33 min 70 min Dnase i handle choloroform / phenol extraction gdna-free Rna 70 min +RT Rt by m-mlv -RT gdna-free cdna gdna-free cdna qrt PCR result compared with the template before and after reverse transcription with gdna clear-up. Sample: Rat A7r5 anchorage-dependent cell (5 x 10 4) before and after reverse transcription with gdna clear-up. Target Gene: rat β-actin. Result: FastLine Cell cdna Kit could totally clean-up gdna. 24

29 RT-PCR Product Product Name ER Quant Reverse Transcriptase 25 rxn ER Quant Reverse Transcriptase 50 rxn ER Quant Reverse Transcriptase 100 rxn ER TIANScript M-MLV 5000 U (25 rxn) ER TIANScript M-MLV U (100 rxn) KR QuantScript RT Kit 25 rxn KR QuantScript RT Kit 100 rxn KR TIANScript RT Kit 25 rxn KR TIANScript RT Kit 100 rxn KR105 FastLine Cell cdna Kit 50 rxn KR FastQuant RT Kit (With gdnase) 25 rxn KR FastQuant RT Kit (With gdnase) 100 rxn KR113 Quant One Step RT-PCR Kit 50 rxn RT-PCR Product 25

30 Real-time PCR Product Real-time PCR Selection Chart of Real-time PCR Products FP202 RealMasterMix (SYBR Green) Real-time PCR (SYBR Green) FP205 SuperReal PreMix Plus (SYBR Green) SYBR Green Two-Step RT*-PCR FP302 Quant qrt-pcr (SYBR Green) Kit Real-time PCR Products One-Step RT*-PCR FP303 Quant One Step qrt-pcr (SYBR Green) Kit Real-time PCR Product Probe Real-time PCR (Probe) FP203 RealMasterMix (Probe) One-Step RT*-PCR FP304 Quant One Step qrt-pcr (Probe) Kit * RT: Reverse Transcription 26

31 Real-time PCR Product RealMasterMix (SYBR Green) Quantitative, Real-time PCR Using SYBR Green I The RealMasterMix Reagent provides accurate real-time quantification of DNA and cdna targets in an easy-to-handle form. The kit can be used in real-time PCR of genomic DNA targets, and also in two-step real-time RT-PCR of RNA targets following reverse transcription. The fluorescent dye SYBR Green I in the master mix enables the analysis of many different targets without having to synthesize target specific labeled probes. High specificity and sensitivity in PCR are achieved by the use of the hot-start enzyme HotMaster Taq DNA Polymerase together with a specialized PCR buffer. The Kit also contains ROX dye, which allows fluorescence normalization on certain cyclers. The kit has been optimized for use with any real-time cycler, including instruments from Applied Biosystems, Bio-Rad, MJ Research, Corbett, Eppendorf, Roche, and Stratagene. Features FP rxn 20 µl FP rxn 20 µl Kit Components 125 rxn 20 µl 500 rxn 20 µl RealMasterMix(2.5 ) 1.0 ml ml 20 SYBR Solution 150 µl 600 µl Store at -20 C and protect from light. Experimental Example 0.2 High amplification efficiency, high specificity: modified HotStar Taq DNA polymerase forming an enzyme automatic regulation system for high amplification efficiency and high specificity real-time PCR. Specific SYBR Solution: meet the demand of different real-time PCR experiment Specifically Mg 2+ automatic regulating system: make sure that your PCR experiment always has a best concentration of Mg 2+. Fluorescence Cycle Real-time PCR Product qrt-pcr result by using TIANGEN FP202. Sample : Rat muscle cdna (gradient dilute from 0.1 to 100 ng) Primer : Rat Jun gen specific primer. PCR volume: 20 µl 27

32 Real-time PCR Product SuperReal PreMix Plus (SYBR Green) Unique two-component Hot-start Taq DNA polymerases for Quantitative, Real-time PCR Using SYBR Green I SuperReal PreMix Plus (SYBR Green) is designed exclusively for SYBR GreenⅠ-based real-time PCR. The kit is premixed with optimal SYBR Green I and 2x SuperReal PreMix Plus which provides easy preparation of reaction solutions. SuperReal PreMix Plus uses unique two-component hot-start Taq DNA polymerases (Chemically modified Taq DNA polymerase and antibody modified Taq DNA polymerase) and special optimal buffer system. This ensures fast, quantitative and sensitive real-time PCR. Features FP FP FP rxn x 20 µl 500 rxn x 20 µl 5000 rxn x 20 µl Components FP FP FP x SuperReal PreMix Plus (with SYBR Green Ⅰ) 50 ROX Reference Dye RNase-free ddh2o 1.25 ml 4 x1.25 ml 10x4x1.25 ml 250 µl 2 x 1ml 1 ml 5 x 1 ml 10 x1 ml 10 x 5 x 1 ml Store at -20 C and in dark condition. Experimental Example SuperReal PreMix Plus uses a unique dual-component hot start polymerase (DNA chemically modified HotStar Taq DNA polymerase and antibody modified), forming an enzyme automatic regulation system, with the carefully optimized buffer system, with high amplification efficiency, high specificity and broad credible range characteristics. Buffer system in the product has balanced the K+ and NH4+ proportion, added a unique H-Bond factor, which can adjust hydrogen bonding of reaction system, making a more rigorous primer and template annealing condition, enhanced reaction specificity, and a better repeatability. To prepare the PCR reaction system, only need to add template, primers, and ddh2o to carry out the Real Time PCR reaction, which is simple and convenient. With ROX Reference Dye, it can eliminate background signal and the correct fluorescence signal between tubes. Reaction system without H-bond Unspecific amplification Reaction system with H-bond Real-time PCR Product Result shows that reaction system containing H-bond could reduce the non-specific amplification effectively dual enzymes system single enzymes dual hot-start enzymes system single enzyme system In most of the reaction systems, dual enzymes system provides a higher sensitive and specific qpcr amplification than the traditional single enzyme system. 28

33 Real-time PCR Product RealMasterMix (Probe) Quantitative, Real-time PCR Using Sequence-Specific Probes RealMasterMix (Probe) Kits provide highly accurate and sensitive detection on any real-time cycler through the combination of the hot-start enzyme HotMaster Taq DNA Polymerase and a unique PCR buffer system. The Kit contains an optimized, ready-to-use master mix for highly sensitive and accurate real-time PCR quantification of genomic DNA or cdna targets using sequence-specific probes. The kit is designed for use with all types of sequence-specific probe, including TaqMan and other dual-labeled probes, FRET probes, and Molecular Beacons. RealMasterMix (Probe) supplied, ready-to-use master mix contains HotMaster Taq DNA Polymerase, ROX passive reference dye, and a dntp mix in an optimized buffer. FP FP rxn 50 µl 200 rxn 50 µl Kit Components 50 rxn 50 µl 200 rxn 50 µl RealMasterMix (2.5 ) 20 Probe Enhancer Solution 1.0 ml 150 µl ml 600 µl Store at -20 C and in dark condition. Experimental Example Features Unique Taq DNA Polymerase: The Kit contains a unique hot-start HotMaster Taq DNA Polymerase. Mg 2+ Adjusting: The Buffer is formulated to adjust the Mg 2+ concentration automatically. Unique Buffer: The kits can be used of any sequence-specific probe on any realtime cycler. Standard Cycle qrt-pcr result by using TIANGEN RealMasterMix (probe) (FP203). Sample : Human total genomic DNA (gradient dilute from 0.1 to 100 ng) Target gene : Sex related gene SRY 29 Real-time PCR Product Amplification Cycle

34 Real-time PCR Product Quant One Step qrt-pcr (SYBR Green) Kit Quantitative, Real-time One-Step RT-PCR Using SYBR Green I Quant One Step qrt-pcr (SYBR Green) Kit provides highly specific SYBR Green detection on any real-time cycler through the combination of a unique RT- PCR buffer system. For convenience, the master mix can be stored at 2-8 C. The kit is also supplied with an optimized RT mix (storage at -20 C) for efficient and sensitive reverse transcription over a wide range of RNA template amounts. FP rxn 50 µl Components 50 rxn 50 µl 2 Quant One Step SYBR RT-PCR Master Mix* 1.3 ml HotMaster Taq Polymerase (2.5 U/µl) 130 µl Quant Reverse Transcriptase (for one step) 30 µl RNase-free ddh 2 O 1.8 ml * Contains dntp Mixture, Mg 2+, SYBR Green I and ROX Reference Dye Store at -20 C and protect from light. Experimental Example Real-time PCR Product Amplification curve Melting curve qrt-pcr result by using TIANGEN Quant One Step qrt- PCR (SYBR Green) Kit (FP303-01) Sample : Human total RNA (gradient dilute from to 100 ng) Target gene : β-actin 30

35 Real-time PCR Product Quant One Step qrt-pcr (Probe) Kit Quantitative, Real-time One-Step RT-PCR Using Sequence-Specific Probes The Quant One Step qrt-pcr (Probe) Kit is supplied with a master mix which contains HotMaster Taq DNA Polymerase, ROX reference dye, and a dntp mixture in an optimized buffer. For convenience, the master mix can be stored at 2-8 C. The kit is also supplied with an optimized RT mix (storage at -20 C) for efficient and sensitive reverse transcription over a wide range of RNA template amounts. Features: Quant One Step qrt-pcr (Probe) Kit provides rapid real-time quantification of RNA targets in an easy-to-handle format and allows both reverse transcription and gene amplification to take place in a single tube. The reaction system is highly sensitive by real-time detection of the PCR products and does not need electrophoresis analysis after PCR, which is suitable for detecting minimal amount of RNA, specially for Virus RNA detection. The high efficient and high fidelity reverse transcriptase and hot start DNA polymerase inside the kit which make a stable and repeatable data. FP rxn 50 µl Components 50 rxn 50 µl 2 Quant One Step Probe qrt-pcr Master Mix* 1.3 ml HotMaster Taq Polymerase (2.5 U/µl) 130 µl Quant Reverse Transcriptase (for one step) 40 µl RNase-free ddh 2 O 1.8 ml * Contains dntp Mixture, One step Probe qrt-pcr buffer and ROX Reference Dye Store at -20 C and protect from light. Experimental Example Amplification curve Real-time PCR Product Standard curve qrt-pcr result by using TIANGEN Quant One Step qrt- PCR (Probe) (FP304). Sample : Human total RNA (gradient dilute from 0.01 to 100 ng) Target gene : HS-UBC gene 31

36 Real-time PCR Product Quant qrt-pcr (SYBR Green) Kit Quantitative, Two-Step RT-PCR Using SYBR Green I Quant qrt-pcr (SYBR Green) Kit provides highly specific SYBR Green detection on any real-time cycler through the combination of HotMaster Taq DNA Polymerase and a unique PCR buffer system. The ready-to-use master mix contains SYBR Green I dye, ROX reference dye, HotMaster Taq DNA Polymerase and a dntp mixture in an optimized buffer. Features The Kit consists of two components: A, Quant First-Strand cdna RT Kit (KR103) and B, RealMasterMix (SYBR Green) (FP202). FP rxn 50 µl FP rxn 50 µl Components 50 rxn 50 µl 200 rxn 50 µl Quant Reverse Transcriptase 25 µl 2 50 µl Oligo(dT) 15 (10 µm) 60 µl 240 µl Random (10 µm) 60 µl 240 µl 10 RT Mix 50 µl 200 µl RNase-free ddh 2 O 1 ml 2 1 ml dntp (2.5 mm each) 60 µl 240 µl RealMasterMix (2.5 ) 1.0 ml ml 20 SYBR Solution 150 µl 600 µl Store at -20 C and protect from light. Experimental Example Real-time PCR Product Amplification curve Standard curve qrt-pcr result by using TIANGEN Quant qrt-pcr (SYBR Green) (FP302) Sample : Human Jurkat Cell total RNA (gradient dilute from 0.01 to 100 ng) Target gene : β-actin 32

37 Real-time PCR Product Product Name FP RealMasterMix (SYBR Green) 125 rxn 20 µl FP RealMasterMix (SYBR Green) 500 rxn 20 µl FP SuperReal PreMix Plus(SYBR Green) 125 rxn x 20 µl FP SuperReal PreMix Plus(SYBR Green) 500 rxn x 20 µl FP SuperReal PreMix Plus(SYBR Green) 5000 rxn x 20 µl FP RealMasterMix (Probe) 50 rxn 50 µl FP RealMasterMix (Probe) 200 rxn 50 µl FP Quant qrt-pcr (SYBR Green) Kit 50 rxn 50 µl FP Quant qrt-pcr (SYBR Green) Kit 200 rxn 50 µl FP Quant One Step qrt-pcr (SYBR Green) Kit 50 rxn 50 µl FP Quant One Step qrt-pcr (Probe) Kit 50 rxn 50 µl Real-time PCR Product 33

38 mirna Analysis Product mircute mirna First-strand cdna Synthesis Kit Reverse Transcription of mirna from 20 pg to 2 µg RNA per Reaction mircute mirna First-strand cdna Synthesis Kit adds Poly(A) to the 3 -terminal of mirna and synthesize the first-strand cdna based on Poly(A) modified mirna through oligo(dt)-universal tag primed reverse transcription. The kit contains all the necessary reagents in the process of adding Poly(A) and reverse transcription. The kit can provide high efficient Poly(A) modification and reverse transcription and efficiently synthesize the first-strand cdna from 20 pg-2 μg total RNA. Principle of mircute mirna cdna synthesis KR KR rxn 50 rxn Kit Components 25 rxn 50 rxn E.coli Poly(A) Polymerase (5U/ µl) 14 µl 28 µl 10 x Poly(A) Polymerase Buffer 60 µl 120 µl 5 x ratp solution 120 µl 240 µl 10 x RT primer (10 µm) 60 µl 120 µl 10 x RT Buffer (10 µm) 70 µl 150 µl dntp (0.5 mm each) 30 µl 60 µl Quant RTase (1U/ µl) 15 µl 30 µl RNasin (40 U/µl) 30 µl 60 µl RNase-free ddh 2 O 1 ml 1 ml Store at -20 C. mirna Analysis Product Important Note Must be used along with mircute mirna qpcr Detection Kit (FP401) 34

39 mirna Analysis Product mircute mirna qpcr Detection Kit (SYBR Green) Detect single base difference between different mirnas in total RNA at the level of pg FP rxn X50 µl mircute mirna qpcr Detection Kit (SYBR Green) is a two-component system which contains 2 x mircute mirna premix and reverse primer. The kit is designed exclusively for detection of mirna in SYBR Green-based real-time PCR. The kit is a new type of qpcr detection system which is premixed with optimal dntps, enhancer, and stabilizers etc. The Taq DNA polymerase uses new antibody modified hotstart technology to ensure high specifility and sensitivity. At the same time, ROX is added into the premix as internal control to make qpcr more accuracte and reliable. Kit Components 50 rxn 2 mircute mirna premix (with SYBR, ROX) 50 ROX Reverse Primer (10 µm) 1.35 ml 250 µl 55 µl Store at 4 C. Do not freeze-thraw repeatly. Experimental Example Features High throughout: detect multiple mirnas from single cdna synthesis reaction. Spedificity: Use antibody modified hotstart PCR polymerase technology and optimal reaction system to avoid non-specific amplification. High resolution: Dectect single base difference of mirna. High sensitivity: Detect specific mirna from total RNA at the level of pg. Important Note Hsa-let-7a family gene preforms a high sequence homology with only several or single nucleotide difference between each other. Real-time PCR analysis result of 5 hsa-let-7 mirna which belongs to Has-let-7a shows that, TIANGEN mirnacute mirna qpcr Detection Kit (SYBR Green) (FP401), could detect single nucleotide difference between them. Amplification curve Melting curve Standard curve qrt-pcr result by using TIANGEN mirnacute mirna qpcr Detection Kit (SYBR Green)(FP401) Sample : Rat liver mirna purified by TIANGEN mircute mirna Isolation Kit (DP501); cdna synthesis by TIANGEN mircute mirna Frist-strand cdna Synthesis Kit (KR201) (gradient dilute from 0.02 to 2000 ng) Target mirna : mir mirna Analysis Product Should be used along with mircute mirna First-strand cdna Synthesis Kit (KR201)

40 DNA Molecular Weight Marker 精准 清晰的DNA分子量Marker 36

41 DNA Molecular Weight Marker Product Name MD DNA Marker I MD DNA Marker I MD DNA Marker II MD DNA Marker II MD DNA Marker III MD DNA Marker III MD DNA Marker IV MD DNA Marker IV MD DNA Marker V MD DNA Marker V MD DNA Marker VII MD DNA Marker VII MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD bp DNA Ladder MD kb DNA Ladder MD kb DNA Ladder MD kb Plus DNA Ladder MD kb Plus DNA Ladder MD D2000 MD D2000 MD D15000 MD D15000 MD D MD D MD λ DNA/Hind III MD λ DNA/Hind III MD pbr322 DNA/Msp I 20 preps MD pbr322 DNA/Msp I 100 preps MD puc18 DNA/Msp I 20 preps MD puc18 DNA/Msp I 100 preps Directly load 3-6 μl of this product on the agarose gel (load 1μl per 1 mm gel lane). To load 6 μl of the product will provide 50 ng per DNA fragment with one intense fragment of 100 ng. 37

42 Genomic DNA Purification Products Product Name DP TIANamp Bacteria DNA Kit DP TIANamp Genomic DNA Kit DP TIANamp Genomic DNA Kit DP Plant Genomic DNA Kit DP Plant Genomic DNA Kit DP TIANamp Yeast DNA Kit DP TIANamp N96 Blood DNA Kit 4 96 preps DP315 TIANamp Virus DNA/RNA Kit DP316 TIANamp Micro DNA Kit DP TIANamp Blood DNA Kit(0.1-1 ml) DP TIANamp Blood DNA Kit (0.1-1 ml) DP RelaxGene Blood DNA System ( ml) 50 ml DP RelaxGene Blood DNA System ( ml) 200 ml DP DNAsecure Plant DNA Kit DP DNAsecure Plant DNA Kit DP DNAquick Plant System DP DNAquick Plant System DP TIANamp Swab DNA Kit DP TIANamp Swab DNA Kit DP TIANamp Feedstuff Animal DNA Kit DP TIANamp Feedstuff Animal DNA Kit DP TIANamp Marine Animal DNA Kit DP TIANamp Marine Animal DNA Kit DP preps DNA Extraction Kit for Food DP Magnetic Genomic DNA Kit DP TIANamp Stool DNA Kit DP TIANquick FFPE DNA Kit DP TIANamp FFPE DNA Kit DP TIANamp Blood DNA Midi Kit 10 preps DP TIANamp Blood DNA Maxi Kit 10 preps KG TIANcombi DNA Lyse&Amp PCR Kit KG TIANcombi DNA Lyse&Amp PCR Kit Individual Buffers and Accessories 38 Product Name RT Red Cell Lysis Buffer 100 ml RT Red Cell Lysis Buffer 250 ml RT401 Lysozyme (50 mg/ml) 1 ml RT Proteinase K (20 mg/ml) 500 μl RT Proteinase K (20 mg/ml) 5 ml RT RNaseA (10 mg/ml) 200 μl RT RNaseA (10 mg/ml) 1 ml RT RNaseA (100 mg/ml) 400 μl RT Lyticase (10 U/µl) 3000 U

43 RNA Purification Products Product Name DP315 TIANamp Virus DNA / RNA Kit DP315-R TIANamp Virus RNA Kit DP TRNzol Reagent 20 ml DP TRNzol Reagent 100 ml DP RNAstore Reagent 20 ml DP RNAstore Reagent 100 ml DP409 RNAsafe (RNase Inhibitor) 1 ml DP412 RNAclean Kit 20 preps DP418 RNasin (RNase Inhibitor) 30 µl DP419 RNAsimple Total RNA Kit DP420 RNAprep pure Kit (For Micro Sample) DP421 TRNzol-A+ Reagent 100 ml DP422 TIANamp DNA-RNA Isolation Kit DP423 TIANamp DNA-RNA-Protein Isolation Kit DP430 RNAprep pure Kit (For Cell/Bacteria) DP431 RNAprep pure Kit (For Tissue) DP432 RNAprep pure Kit (For Plant) DP433 RNAprep pure Kit (For Blood) DP439 RNAprep pure FFPE Kit DP RNALock Reagent 20 ml DP RNALock Reagent 100 ml DP501 mircute mirna Isolation Kit Plasmid DNA Purification Products Product Name DP TIANprep Mini Plasmid Kit DP TIANprep Mini Plasmid Kit DP TIANpure Mini Plasmid Kit DP TIANprep Midi Plasmid Kit DP TIANpure Midi Plasmid Kit DP TIANprep Yeast Plasmid DNA Kit DP TIANprep Highpure N96 Plasmid Kits 4x96 wells deepwell plates DP TIANprep Highpure N96 Plasmid Kits 24x96 wells deepwell plates DP TIANprep Rapid N96 Plasmid Kits 4 96 preps DP TIANprep Rapid N96 Plasmid Kits preps DP116 HighPure Maxi Plasmid Kit 10 preps DP117 EndoFree Maxi Plasmid Kit 10 preps 39

44 PCR Cleanup & Gel Extraction Products 40 Product Name DP TIANquick Midi Purification Kit DP TIANquick Midi Purification Kit DP TIANquick Maxi Purification Kit DP TIANgel Midi Purification Kit DP TIANgel Midi Purification Kit DP TIANgel Maxi Purification Kit DP TIANquick Oligo Purification Kit 20 preps DP TIANquick N96 Purification Kit 4 96 preps DP TIANquick N96 Purification Kit preps DP Universal DNA Purification Kit DP Universal DNA Purification Kit

45 Cloning Products pgm-t Vector Map pgm-t Vector Cloning Site Product Name VT pgm-t Ligation Kit 20 µl VT pgm-t Ligation Kit 60 µl 41

46 pzeroback/blunt Vector Map pzeroback/blunt Vector Cloning Site 42 Product Name VT ZeroBack Fast Ligation Kit 20 µl VT ZeroBack Fast Ligation Kit 10 µl