Development of a new GMP TAQ polymerase for improved PCR performance. Marie-Claire Beckers, Peter Haima
|
|
|
- Ambrose Blair
- 5 years ago
- Views:
Transcription
1 Development of a new GMP TAQ polymerase for improved PCR performance Marie-Claire Beckers, Peter Haima
2 Contents Context: Eurogentec GMP manufacturing of IVD components PCR performance issues Novel concept: Hot Diamond Taq Mode of action Proof of principle Conclusions
3 Our Activities PRODUCTS & SERVICES for RESEARCH Manufacturing of custom oligos, reagents & enzymes for (Q)PCR, antibodies, proteins, peptides. CMO SOLUTIONS DIAGNOSTICS GMP compliant ISO13485 certified manufacturer of oligonucleotides, Taq polymerases, peptides and antibodies for R&D and commercial IVD products. CMO SOLUTIONS PHARMA cgmp compliant manufacturer of biopharmaceuticals from bacterial/yeast sources:
4 Fremont San Diego Five Manufacturing Sites Belgium Japan
5 CMO for IVD industry EGT is a leading European manufacturer of research grade oligonucleotides since IVD GMP oligos to global IVD industry since 04. GMP manufacturing and ISO certification for IVD oligos in Europe ( 08), Japan ( 09) and USA( 10). GMP (pharma) production of Taq Polymerases in 2009.
6 Polymerase Chain Reaction > fold amplification of a nucleic acid sequence. Revolutionized molecular research and IVD because of unparalleled sensitivity and specificity (HIV, H1N1). Inherent advantages & disadvantages.
7 PCR issues Enzymes contain E. coli & fungal DNA which can lead to false positives. Contamination and off target amplification, which can lead to false positives. Primer-dimer formation reduces yield of specific amplicon, which can lead to false negatives. Royalty stacking is a concern for IVD companies.
8 GMP TAQ Polymerases Diamond Taq
9 Diamond Taq recombinant non hot-start Manufacturing in GMP-Pharma (QMS/clean rooms). Fully compliant ISO & GMP/QSR Customized Fill & Finish. Batch record & CoA. DNA ultralow (<0.01 fg/u). Bioburden ultralow (typically 0; < 10 CFU/ml). Exceptional lot to lot reproducibility.
10 PCR issues Enzymes contain E. coli & fungal DNA which can lead to false positives. Contamination and off target amplification, which can lead to false positives. Primer-dimer formation reduces yield of specific amplicon, which can lead to false negatives. Royalty stacking is a concern for IVD companies.
11 Hot start PCR Most effective strategy to reduce nonspecific amplification during low temperature set-up. Wax-embedded PCR reagents. Modified DNA polymerase, inactive (< 74 C) prior to heat shock (up to C) Hot-start dntps. PCR Hot Start by magnesium sequestration.
12 Hot start DNA Polymerases Activity modified with blocking antibody. Disadvantage: antibody from hybridomas can contaminate PCR with mammalian DNA; enzymes also tend to be leaky. Activity modified chemically. Disadvantage: requires long activation (10 mins), could cause heat-damage to DNA samples.
13 New Hot Start Concept NEW CONCEPT HOT Diamond Taq Not through chemical modification. Not through blocking antibody. Enzyme shows no activity at room temperature. No heat shock is required; 100% enzyme activity appears in 1 st PCR cycle. Very high yield of specific PCR product.
14 HOT Diamond Taq Proprietary agent prevents non-specific polymerization; effectively prevents primerdimer formation, increases specific PCR yield. Hot Diamond TAQ buffer is designed to maximize effect of the proprietary agent. Hot Diamond TAQ protocol is compatible with all existing hot-start protocols.
15 HOT Diamond Taq Universal concept, Hot Diamond Taq can be used to amplify AT and GC rich regions, long and difficult templates. Production under GMP conditions provides enzyme with ultra low bioburden and E.coli/fungal DNA content. Can be used for demanding Research and IVD applications (sensitivity, specificity, GMP). Universal concept, can also be used to improve performance other (hot-start) enzymes.
16 Polymerisation test Elongation Incubation at 25 C HGS GMP TOP HGS GMP TOP Elongated hybrid Oligo lc hybrid Oligo l Without activation Oligo c With activation
17 Polymerisation test Elongation Incubation at 25 C HGS GMP TOP HGS GMP TOP Without With activation activation 1: Oligoc 2: Oligol 3: Hybrid Oligoc-Oligol 4: Double-strand Oligol 5: Hot Start : 6h30 at 25 C 6: Hot Diamond : 6h30 at 25 C 7: Hot Diamond : 6h30 at 25 C 8: Hot Start : 10min at 95 C and 6h30 at 25 C 9: Hot Diamond : 10min at 95 C and 6h30 at 25 C 10: Hot Diamond : 10min at 95 C and 6h30 at 25 C
18 Polymerisation test Elongation Incubation at 40 C : Hybrid Oligoc-Oligol 2: Double-strand Oligol 3: Hot Start 4h at 40 C 4: Hot Diamond 4h at 40 C 5: Hot Diamond 4h at 40 C 7: Hot Start C/4h at 40 C 8: Hot Diamond C/4h at 40 C 9: Hot Diamond C/4h at 40 C Without activation With activation
19 Polymerisation test Elongation Incubation at 60 C HGS GMP TOP HGS GMP TOP Without activation Without activation With activation With activation 1: Oligoc 2: Oligol 3: Hybrid Oligoc-Oligol 4: Double-strand Oligol 5: Hot Start 4h at 60 C 6: Hot Diamond 4h at 60 C 7: Hot Diamond 4h at 60 C 8: Hot Start C/4h 60 C 9: Hot Diamond C/4h 60 C 10: Hot Diamond C/4h 60 C
20 Performance Test Lambda DNA Amplification SmartLadder 2.Lambda DNA 100 pg 3. Lambda DNA 10 pg 4. Lambda DNA 1 pg 5. Lambda DNA 0.1 pg Thermal cycling conditions: 94 C 3' 25 cycle 94 C 30'' 60 C 1' 72 C 2' 72 7' 4 C end temperature
21 Hot Diamond Taq 1: SmartLadder 2: NTC 3: 0,1ng DNA 4: NTC 5: 0,1ng DNA 6: SmartLadder 7: NTC, 8: 0,1ng DNA 9: NTC 10: 0,1ng DNA 11: SmartLadder Non-Hot Start Taq 1: SmartLadder 2: NTC 3: 0,1ng DNA 4: NTC 5: 0,1ng DNA 6: SmartLadder 7: NTC 8: 0,1ng DNA 9: NTC 10: 0,1ng DNA 11: SmartLadder AT rich gene from H.Pylori GC rich gene from S. Coelicolor Setup Incubation: 4h at 25 C Thermal cycling conditions: 3 min 95 C 40 cycles 30 sec at 95 C 30 sec at 50 C for AT rich sequence or 30 sec at 60 C for GC rich sequence 1 min at 72 C/ 5 min at 72 C Performance Test Multiple Targets
22 Performance Test Different Taq Polymerases Numb gene Amplification 1. SmartLadder 2. Hot Diamond 3. Competitor A Hotstart Taq 4. Competitor B Hotstart Taq 5. Competitor C Hotstart Taq 6. Competitor D Hotstart Taq 7. Diamond Taq GC rich gene Amplification 8. SmartLadder 9. Hot Diamond 10. Competitor A Hotstart Taq 11. Competitor B Hotstart Taq 12. Competitor C Hotstart Taq 13. Competitor D Hotstart Taq 14. Diamond Taq
23 Performance Test Multiple Targets & Different Taq Polymerases λ DNA /8. SmartLadder 2/9. Hot Diamond Taq 3/10. Competitor A Hotstart Taq 4/11. Competitor B Hotstart Taq 5/12. Competitor C Hotstart Taq 6/13. Competitor D Hotstart Taq 7/14. Diamond Taq Gluco3203 (3.2 Kb) Thermal cycling conditions (Gluco3203): 1 min 94 C First 10 cycles: 10 sec at 94 C 60 sec at 60 C 5 min at 68 C Next 20 cycles: 10 sec at 94 C 60 sec at 60 C 5 min at 68 C + 10sec each cycle 5 min at 72 C 4 C
24 Lambda DNA Amplification Different Times of Heat Activation Hot Diamond Taq Hot Start Taq Thermal cycling conditions: 94 C 10 min, 5 min or 20 seconds 25 cycles 94 C 30'' 60 C 1' 72 C 2' 72 7' 4 C end temperature 1. SmartLadder 2. λ DNA 5 pg - 20 sec 3.λDNA 1 pg - 20 sec 4. NTC - 20 sec 5. λ DNA 5 pg - 5 min 6. λ DNA 1 pg - 5 min 7. NTC - 5 min 8. λ DNA 5 pg - 10 min 9. λ DNA 1 pg - 10 min 10. NTC - 10 min
25 Real-Time PCR Performance Probe Assay : TATA box binding protein gene Relative low expression level Hot Diamond Taq Hot Start Taq Samples Hot Start Hot Diamond Ct Ct 100ng cdna 30,07 30,15 50ng cdna 31,4 31,28 25ng cdna 32,42 32,28 12,5ng cdna 33,46 33,44 6,25ng cdna 34,78 34,23 3,13ng cdna 34,9 36,58 NTC x x Thermal cycling conditions: 2 min at 50 C 10min at 95 C 40 cycles: 15sec at 95 C 1 min at 60 C
26 Quantification of residual gdna from E. coli qpcr qpcr qpcr
27 Quantification of residual gdna from E. coli Hot Diamond Taq Hot Start other fg DNA/ Taq unit Genome copies/ Taq unit
28 Hot Diamond TAQ concept applied to other enzymes Numb gene 1. gdna 10 ng 2. gdna 1 ng 3. gdna 0.1 ng 4. NTC HS TAQ 1 Modified HS TAQ 2 Modified PCR conditions 3 min 95 C 35 cycles: 30 sec at 95 C 10 sec at 60 C 30 sec at 72 C 5 min at 72 C 4 C
29 Conclusions Hot Diamond Taq demonstrated equal to better performance in direct comparison with other HotStart DNA polymerases in PCR and qpcr applications without the «Hot start» issues. Complete enzyme activation provides maximal enzymatic activity during PCR to generate higher yields. Universal concept, can also be used to improve performance other (hot-start) enzymes.
30 Conclusions Hot Diamond Taq improves PCR efficiency and specificity to amplify AT and GC rich regions, long and difficult templates. Hot Diamond Production under strict GMP conditions provides an ultrapure bioburden-free enzyme and extremely low in residual E.coli DNA. Hot Diamond Taq is highly suited for all IVD and demanding research applications and bacterial and fungal PCR assays. (sensitivity, specificity, GMP). Hot Diamond Taq may also be suitable for isothermal amplification.
31 Arigatou gozaimasu Thank you very much!
USB HotStart-IT. for increased specificity and consistent results. PCR, qpcr and qrt-pcr
USB HotStart-IT for increased specificity and consistent results PCR, qpcr and qrt-pcr USB PCR Reagents Choose USB HotStart-IT products for increased specificity and consistent results. Long and Accurate
P HENIX. PHENIX PCR Enzyme Guide Tools For Life Science Discovery RESEARCH PRODUCTS
PHENIX PCR Enzyme Guide PHENIX offers a broad line of premium quality PCR Enzymes. This PCR Enzyme Guide will help simplify your polymerase selection process. Each DNA Polymerase has different characteristics
Functional Genomics Research Stream. Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update
Functional Genomics Research Stream Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update Updates Alternate Lab Meeting Fridays 11:30-1:00 WEL 4.224 Welcome to attend either one Lab Log thanks
Uses: The Methylamp MS-qPCR Fast Kit is very suitable for quantitative methylation-specific PCR in a fast format using very minute amounts of DNA.
Methylamp MS-qPCR Fast Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE Uses: The Methylamp MS-qPCR Fast Kit is very suitable for quantitative methylation-specific PCR in a fast format
DNA Amplification RNA Amplification Real-Time PCR Customized PCR
DNA Amplification RNA Amplification Real-Time PCR Customized PCR Selection Guide Overview AccuPower PreMix series is an innovative PCR master mix product range designed to make your PCR, reverse transcription
THUNDERBIRD SYBR qpcr Mix
Instruction manual THUNDERBIRD SYBR qpcr Mix 1304 A4251K THUNDERBIRD SYBR qpcr Mix QPS-201T 1 ml x 1 QPS-201 1.67 ml x 3 Contents [1] Introduction [2] Components [3] Primer design [4] Template DNA [5]
Executive Summary. clinical supply services
clinical supply services case study Development and NDA-level validation of quantitative polymerase chain reaction (qpcr) procedure for detection and quantification of residual E.coli genomic DNA Executive
AccuPower PCR PreMix 73. AccuPower Taq PCR PreMix 77. AccuPower PCR PreMix (with UDG) 79. AccuPower HotStart PCR PreMix 81
PCR PreMix 73 AccuPower Taq PCR PreMix 77 AccuPower PCR PreMix (with UDG) 79 AccuPower HotStart PCR PreMix 81 AccuPower PyroHotStart Taq PCR PreMix 84 AccuPower HotStart PCR PreMix (with UDG) 87 AccuPower
SYBR Premix Ex Taq II (Tli RNaseH Plus), ROX plus
Cat. # RR82LR For Research Use SYBR Premix Ex Taq II (Tli RNaseH Plus), ROX plus Product Manual The long-term storage temperature of this product has been changed to -20 since Lot. A1901A. See section
Critical Factors For Successful Real-Time RT-PCR
Critical Factors For Successful Real-Time RT-PCR Andreas Missel, PhD Senior Scientist R&D Dept. Modification/Amplification QIAGEN Annealing Specific product High yield High sensitivity Nonspecific product
SYBR Premix Ex Taq II (Tli RNaseH Plus), Bulk
Cat. # RR820L For Research Use SYBR Premix Ex Taq II (Tli RNaseH Plus), Bulk Product Manual The long-term storage temperature of this product has been changed to -20 since Lot. AK9104. See section IV.
Platinum II Taq Hot-Start DNA Polymerase for high-throughput PCR
WHITE PAPER Platinum II Taq Hot-Start DNA Polymerase Platinum II Taq Hot-Start DNA Polymerase for high-throughput PCR Abstract The advances in thermal cycler technology permit a substantial increase in
SunScript TM One Step RT-qPCR Kit
INDEX Ordering Information...3 Kit Contents...3 Shipping and Storage...3 Handling...3 Quality Control...3 Reagents and Equipment to be Supplied by the User...3 Description...4 Protocol...4 Troubleshooting
GreenMasterMix (2X) b i o s c i e n c e. G E N A X X O N b i o s c i e n c e. High ROX (500nM)
G:\products\productflyer\pcr\polymerasen\hotstart\manu_m3052_green_en.docx GreenMasterMix (2) High RO (500nM) qpcr master mix with fluorescence dye and passive reference dye Contact & Technical support
TB Green Premix Ex Taq (Tli RNaseH Plus)
Cat. # RR420A For Research Use TB Green Premix Ex Taq (Tli RNaseH Plus) Product Manual The long-term storage temperature of this product has been changed to -20 since Lot. #AK8101. See section V. Storage.
PCR KIT/REAGENTS/BUFFERS/PRIMERS
PCR KIT/REAGENTS/BUFFERS/PRIMERS 114330 DNA Amplification Kit DNA amplification kit is suitable for amplification of DNA size about 100bp to 5kb. It can be also used to RAPD PCR. This kit contains all
AmpliTaq 360 DNA Polymerase
Product Bulletin AmpliTaq 360 AmpliTaq 360 360 Coverage for a Full Range of Targets AmpliTaq 360 Benefits Optimized for the broadest range of targets (
Cat. # RR430S RR430A. For Research Use. SYBR Fast qpcr Mix. Product Manual. v201610da
For Research Use SYBR Fast qpcr Mix Product Manual Table of Contents I. Description... 3 II. Principle... 4 III. Components... 5 IV. Storage... 5 V. Features... 6 VI. Precautions... 6 VII. Protocol...
Table of Contents. I. Description...2. Components...2. Storage...2. Features...2. V. General Composition of PCR Reaction Mixture...
Table of Contents I. Description...2 II. Components...2 III. Storage...2 IV. Features...2 V. General Composition of PCR Reaction Mixture...5 VI. PCR Conditions...5 VII. Optimization of Parameters...6 VIII.
Technical Review. Real time PCR
Technical Review Real time PCR Normal PCR: Analyze with agarose gel Normal PCR vs Real time PCR Real-time PCR, also known as quantitative PCR (qpcr) or kinetic PCR Key feature: Used to amplify and simultaneously
SpeedSTAR HS DNA Polymerase
Cat. # RR070A For Research Use SpeedSTAR HS DNA Polymerase Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage... 3 IV. Supplied buffers... 3 V. General reaction mixture...
TB Green Premix Ex Taq II (Tli RNaseH Plus)
Cat. # RR820A For Research Use TB Green Premix Ex Taq II (Tli RNaseH Plus) Product Manual The long-term storage temperature of this product has been changed to -20 since Lot. #AGY1013N. See section IV.
Reverse Transcription & RT-PCR
Creating Gene Expression Solutions Reverse Transcription & RT-PCR Reverse transcription, a process that involves a reverse transcriptase (RTase) which uses RNA as the template to make complementary DNA
SunScript One Step RT-PCR Kit
SunScript ONE STEP R T-PCR KIT HANDBOOK SunScript One Step RT-PCR Kit INDEX Legal... 4 Intended use... 4 Kit contents... 5 Shipping and storage... 5 Handling... 6 Quality control... 6 Reagents and equipment...
Roche Molecular Biochemicals Technical Note No. LC 9/2000
Roche Molecular Biochemicals Technical Note No. LC 9/2000 LightCycler Optimization Strategy Introduction Purpose of this Note Table of Contents The LightCycler system provides different detection formats
MOLECULAR SOLUTIONS FOR BIOTECHNOLOGY
MOLECULAR SOLUTIONS FOR BIOTECHNOLOGY OUR PCR TECHNOLOGY OUR CAPABILITIES Over 20 years of molecular biology research has given Bioline scientists a deep understanding of the profound influence that buffer
Premix Ex Taq (Probe qpcr)
For Research Use Premix Ex Taq (Probe qpcr) Product Manual Table of Contents I. Description... 3 II. Principle... 4 III. Components... 5 IV. Materials Required but not Provided... 5 V. Storage... 5 VI.
SYBR Premix DimerEraser (Perfect Real Time)
Cat. # RR091A or Research Use SYBR Premix DimerEraser (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Principle... 3 III. Components... 4 IV. Storage... 5 V. eatures... 5 VI.
PrimeSTAR Max DNA Polymerase
Cat. # R045A For Research Use PrimeSTAR Max DNA Polymerase Product Manual Table of Contents I. Description...3 II. III. IV. Components...3 Storage...3 General Composition of PCR Reaction Mixture...3 V.
qpcr Quantitative PCR or Real-time PCR Gives a measurement of PCR product at end of each cycle real time
qpcr qpcr Quantitative PCR or Real-time PCR Gives a measurement of PCR product at end of each cycle real time Differs from endpoint PCR gel on last cycle Used to determines relative amount of template
KAPA HiFi HotStart ReadyMix PCR Kit
KAPA HiFi HotStart ReadyMix PCR Kit KR0370 v10.19 This provides product information and a detailed protocol for the KAPA HiFi HotStart ReadyMix PCR Kit. This document applies to the following kits: 07958919001,
SYBR Green Realtime PCR Master Mix -Plus-
Instruction manual SYBR Green Realtime PCR Master Mix -Plus- 0803 F0925K SYBR Green Realtime PCR Master Mix -Plus- Contents QPK-212 1mLx5 Store at -20 C, protected from light [1] Introduction [2] Components
BEST QUALITY HIGHEST PURITY. Recombinant ENZYMES & PROTEINS
BEST QUALITY HIGHEST PURITY Recombinant ENZYMES & PROTEINS We offer a wide range of highest quality enzymes and proteins for molecular biology including DNA polymerases, reverse transcriptases, DNA ligases,
Frequently Asked Questions
Frequently Asked Questions VeriQuest qpcr Master Mixes VeriQuest Probe qpcr Master Mix 1. What is the VeriQuest Probe qpcr Master Mix? VeriQuest Probe qpcr Master Mix is a ready-to-use mix for real-time
Cat. # RR391A. For Research Use. Probe qpcr Mix. Product Manual. v201610da
Cat. # RR391A For Research Use Probe qpcr Mix Product Manual Table of Contents I. Introduction... 3 II. Principle... 3 III. Components... 5 IV. Materials Required but not Provided... 5 V. Storage... 5
Optimizing a Conventional Polymerase Chain Reaction (PCR) and Primer Design
Optimizing a Conventional Polymerase Chain Reaction (PCR) and Primer Design The Polymerase Chain Reaction (PCR) is a powerful technique used for the amplification of a specific segment of a nucleic acid
EpiQuik Quantitative PCR Fast Kit Base Catalog # P-1029
EpiQuik Quantitative PCR Fast Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE Uses: The EpiQuik Quantitative PCR Fast Kit is designed for quantitative real time analysis of DNA samples
Our new breed is the center of attention.
AMPLIFICATION CELL BIOLOGY CLONING MICROARRAYS NUCLEIC ACID ANALYSIS PROTEIN FUNCTION & ANALYSIS QUANTITATIVE PCR SOFTWARE SOLUTIONS Our new breed is the center of attention. II enzyme for highest fidelity
Guidelines for Developing Robust and Reliable PCR Assays
Guidelines for Developing Robust and Reliable PCR Assays Leta Steffen, PhD Applications Scientist Promega Corporation Outline 1) PCR reaction components What is in the reaction? How does it affect assay
Procomcure Biotech. PCR Reagents
Procomcure Biotech PCR Reagents valid for 2018 VitaTaq DNA Polymerase is a standard Taq DNA polymerase suitable for all common PCR applications like colonypcr, cloning applications, high-throughput PCR
FAQs: PCR Polymerases from Takara Bio
FAQs: PCR Polymerases from Takara Bio Contents: PCR Basics Q1 Q2 Q3 What parameters do I need to consider when designing primers? What is the optimum amount of template to use? Which conditions are particularly
One Step SYBR PrimeScript RT-PCR Kit II (Perfect Real Time)
Cat. # RR086A For Research Use One Step SYBR PrimeScript RT-PCR Kit II Product Manual Table of Contents I. Description...3 II. III. IV. Principle...3 Components...5 Storage...6 V. Features...6 VI. VII.
PlantDirect TM Multiplex PCR System
PlantDirect TM Multiplex PCR System Technical Manual No. 0178 Version 10112010 I Description.. 1 II Applications 2 III Key Features.. 3 IV Shipping and Storage. 3 V Simplified Procedures. 3 VI Detailed
SYBR Green Realtime PCR Master Mix
Instruction manual SYBR Green Realtime PCR Master Mix 0810 F0924K SYBR Green Realtime PCR Master Mix QPK-201T 1 ml x 1 QPK-201 1 ml x 5 Contents [1] Introduction [2] Components [3] Primer design [4] Detection
2x PCR LongNova-RED PCR Master Mix
2x PCR LongNova-RED Components RP85L 100 reactions (50 μl) RP85L-10 1000 reactions (50 μl) 2x PCR LongNova-RED 2 x 1.25 ml 20 x 1.25 ml PCR grade water 2 x 1.5 ml 20 x 1.5 ml Storage & Shiing Storage conditions
The Next Generation in Hot Start PCR - CleanAmp Primers and dntps. Natasha Paul, Ph.D. TriLink BioTechnologies, Inc.
The Next Generation in Hot Start PCR - CleanAmp Primers and dntps Natasha Paul, Ph.D. TriLink BioTechnologies, Inc. The Polymerase Chain Reaction Thermal cycling process that provides ~ 10 6 -fold amplification
High Yield/ Routine Applications Problematic templates, High GC/AT High Fidelity Heat-Activated Long Products High Specificity DHPLC Compatible
Yield/ Routine Applications Problematic templates, GC/AT Fidelity Heat-Activated Long Products Specificity DHPLC Compatible Polymerases Optimised dntp/ Polymerase Mixes Introduction are essential tools
10x Hot-Taq Buffer 1 ml 1 ml x 2ea 1 ml x 10ea. dntp Mix (each 10 mm) None / 0.2 ml None / 0.4 ml None / 0.4 ml x 5ea
HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) CERTIFICATE OF ANALYSIS (1603-V01R03) Contents HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) Cat. No. HT250/HT250N HT500/HT500N HT2500/HT2500N Hot-Taq (2.5 units/μl)
HiPer Real-Time PCR Teaching Kit
HiPer Real-Time PCR Teaching Kit Product Code: HTBM032 Number of experiments that can be performed: 10 Duration of Experiment Protocol: 1.5 hours Storage Instructions: The kit is stable for 12 months from
SYBR Advantage qpcr Premix. User Manual
User Manual SYBR Advantage qpcr Premix User Manual United States/Canada 800.66.566 Asia Pacific +.650.99.7300 Europe +33.(0).3904.6880 Japan +8.(0)77.543.66 Clontech Laboratories, Inc. A Takara Bio Company
2X Q-PCR Master Mix 1 ml x 2 (SYBR, ROX) Storage Aliquot to avoid multiple freeze-thaw cycles Protect from light -20 C for 12 months
www.smobio.com Product Information ExcelTaq series 2X Q-PCR Master Mix (SYBR, ROX) TQ1110 200 RXN 2X Q-PCR Master Mix 1 ml x 2 (SYBR, ROX) Storage Aliquot to avoid multiple freeze-thaw cycles Protect from
Ver. 3(EN) Selection Guide of Bioneer s PCR kits
20151116 Ver. 3(EN) Selection Guide of Bioneer s PCR kits 8-11, Munpyeongseoro, Daedeok-gu, Daejeon, 34302, Republic of Korea Tel: (Korea) 1588-9788 (International) +82-42 - 930-8777 Email: sales@bioneer.com
Polymerase Chain Reaction PCR
1 Description of Module Subject Name Paper Name Module Name/Title Dr. Vijaya Khader Dr. MC Varadaraj 2 1. Objectives 1. To understand principle of 2. Types 3. Applications 2. Lay Out 3 Types of Qualitative
Amplification Products for PCR and RT-PCR
Selection guide Polymerase Hot start Comment UptiTherm DNA pol. no Most economic. Lower error rate than Taq polymerase Available in several formats, master mix including or not dntp, Mg 2+..., in gel format
Brilliant III Ultra-Fast SYBR Green QPCR Master Mix
Brilliant III Ultra-Fast SYBR Green QPCR Master Mix Instruction Manual Catalog #600882 (single kit) #600883 (10-pack kit) Revision D0 Research Use Only. Not for Use in Diagnostic Procedures. 600882-12
Going MULTI How to Easily Achieve High Multiplexing in Real-Time PCR
qpcr Symposium 2005 - Weihenstephan Going MULTI How to Easily Achieve High Multiplexing in Real-Time PCR Dr. Andreas Missel Associate Director Research & Development Overview Introduction Multiplex real-time
QUICK-Clone TM User Manual. cdna
QUICK-Clone TM User Manual cdna PT1150-1 (PR752268) Published 25 May 2007 Table of Contents I. Introduction 3 II. Applications Discussion 4 A. Primer Design 4 B. Setting up the PCR Reaction 4 C. Example
SYBR Premix Ex Taq II (Tli RNaseH Plus), Bulk
Cat. # RR820L For Research Use SYBR Premix Ex Taq II (Tli RNaseH Plus), Bulk Product Manual Table of Contents I. Description... 3 II. Principle... 3 III. Kit Components... 4 IV. Materials Required but
Methods in virus diagnosis PCR techniques
Methods in virus diagnosis PCR techniques 450 MBIO PRACTICAL LESSON 5 Molecular Methods Methods based on the detection of viral genome are also commonly known as molecular methods. It is often said that
TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 2 TQ RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 5
www.smobio.com Product Information ExcelTaq series 2X Q-PCR Master Mix (SYBR, no ROX) TQ1100 200 RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x 2 TQ1101 500 RXN 2X Q-PCR Master Mix (SYBR, no ROX) 1 ml x
Bauer Core Standard Protocol Title: Guidelines for Designing Real Time PCR Experiments Pages: 5 Revision: 1.1 Date: 4/15/04
Bauer Core Standard Protocol Title: Guidelines for Designing Real Time PCR Experiments Pages: 5 Revision: 1.1 Date: 4/15/04 Author(s): Claire Reardon Reviewers: Christian Daly Contact: claire@cgr.harvard.edu
Premix Ex Taq (Probe qpcr)
For Research Use Premix Ex Taq (Probe qpcr) Product Manual Table of Contents I. Description... 3 II. Principle... 4 III. Components... 5 IV. Materials Required but not Provided... 5 V. Storage... 5 VI.
PrimeScript 1st strand cdna Synthesis Kit
Cat. # 6110A For Research Use PrimeScript 1st strand cdna Synthesis Kit Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided... 3 IV. Storage...
PCR Amplifies Targeted Sequence
PCR Amplifies Targeted Sequence Target Sequence Supercoiled DNA Strand DNA Strand Double Helix DNA Strand Chromosome P C R PCR PCR = Polymerase Chain Reaction. A primer directed-extension reaction for
Taq + dntps #EZ U
#EZ1012 500U Taq + dntps Concentration: 5U/μl Contents: Hy-Taq DNA polymerase 100μl 10xPCR Buffer (Mg2+ Plus) 1.25ml dntps(2.5mm each) 1ml 6xLoading Buffer 1ml Store at -20 C Description Hy-Taq 500U +
Accura High-Fidelity Polymerase
Accura High-Fidelity Polymerase FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE Lucigen Corporation 2905 Parmenter St, Middleton, WI 53562 USA Toll Free: (888) 575-9695 (608) 831-9011 FAX: (608)
winter savings one place High Quality Costs Less Look inside to find great deals on Thermo Scientific products for all your molecular biology needs
A quarterly publication containing special offers for significant savings on a variety of molecular biology products CDA winter savings High Quality Costs Less RNAi PCR / qpcr Look inside to find great
2 march 06 Seminar on RT-PCR. About Real-time PCR. Aurélie OLIVIER Université Catholique de Louvain Unité de pharmacologie cellulaire et moléculaire
2 march 06 Seminar on RT-PCR About Real-time PCR Aurélie OLIVIER Université Catholique de Louvain Unité de pharmacologie cellulaire et moléculaire Target DNA PCR Applications: Gene Plasmide, phage Diagnostic
Polymerase Chain Reaction PCR
Polymerase Chain Reaction PCR What is PCR? An in vitro process that detects, identifies, and copies (amplifies) a specific piece of DNA in a biological sample. Discovered by Dr. Kary Mullis in 1983. A
HELINI Hepatitis B virus [HBV] Real-time PCR Kit (Genotype A to H)
![HELINI Hepatitis B virus [HBV] Real-time PCR Kit (Genotype A to H)](/thumbs/87/95094490.jpg "HELINI Hepatitis B virus [HBV] Real-time PCR Kit (Genotype A to H)")
HELINI Hepatitis B virus [HBV] Real-time PCR Kit (Genotype A to H) Quantitative In vitro diagnostics Instruction manual Cat. No: 8001-25/50/100 tests Compatible with: Agilent, Bio-Rad, Applied Bio systems
FastGene Optima. Products for PCR. proof-reading. ReadyMix. cdna SNP. routine PCR. polymerase. optimized blend complex templates. N gene knock out B O
Products for PCR www.nippongenetics.eu efficiency polymerase convenience endpoint PCR incl. loading dye direct PCR incl. dntps engineered enzyme archeal type B polymerase high GC content FastGene tissuebest
Next Generation Polymerase Chain Reaction
Next Generation Polymerase Chain Reaction Developed by Nobel laureate Kary Mullis in the 1980s, Polymerase Chain Reaction (PCR) is a molecular technology that allows fast and in vitro. It has since become
PRODUCT INFORMATION Thermo Scientific Luminaris Probe Low ROX qpcr Master Mix #K0944 For 5000 rxns Lot Expiry Date Store at -20 C in the dark CERTIFICATE OF ANALYSIS The absence of endo-, exodeoxyribonucleases
SpeAmp n cdna Pre-amplification Kits User Guide
SpeAmp n cdna Pre-amplification Kits User Guide Cat # SpeAmp n -10 Cat # SpeAmp n -25 Cat # SpeAmp n -50 To use with our range of pathway-specific primer pools : Cat # Prim nx -10 XXXX Cat # Prim nx -25
Brilliant II Fast SYBR Green QPCR Master Mix
Brilliant II Fast SYBR Green QPCR Master Mix INSTRUCTION MANUAL Catalog #600843 (single kit) #600844 (10-pack kit) Revision B.01 For In Vitro Use Only 600843-12 LIMITED PRODUCT WARRANTY This warranty limits
Detecting Rare Genomic Copies or Events
Detecting Rare Genomic Copies or Events Integrated DNA Technologies Nick Downey, PhD Common Diagnostics Challenges Detecting targets early with accuracy Detecting different strains (e.g., antibiotic resistance)
Brilliant II SYBR Green QPCR Master Mix
Brilliant II SYBR Green QPCR Master Mix INSTRUCTION MANUAL Catalog #600828 (single kit) #600831 (10-pack kit) Revision B.01 For In Vitro Use Only 600828-12 LIMITED PRODUCT WARRANTY This warranty limits
Hy-Fy High Fidelity Mix (x2)
Hy-Fy High Fidelity Mix (x2) #EZ-2021 1ml, 100rxn of 20μl Contents: 2X High Fidelity Mix 1ml Nuclease-free water 1ml Store at -20 C Shelf life: 2 years Description Hy-Fy High Fidelity Mix (x2) is a premixed,
CERTIFICATE OF ANALYSIS. For. Red i-taq DNA Polymerase
CERTIFICATE OF ANALYSIS For Red i-taq DNA Polymerase Version 1 (110804) Catalog i-taq6 Size 500 units For Research Use Only and Not For Human and Animal Therapeutic and Diagnostic Use. Disclaimer: THE
SYBR Real-Time PCR Kit
SYBR Real-Time PCR Kit For Amplification and detection of DNA in Quantitative real-time PCR (qpcr). Catalog No. QPG-040/QPG-041/QPG-042/QPG-043 User Manual Table of Contents Kit Contents and Storage...
Molecular methods for detection of Antibiotic Resistance in environmental matrices: limits, prospects and challenges.
Dr. Angela Cicatelli acicatelli@unisa.it Molecular methods for detection of Antibiotic Resistance in environmental matrices: limits, prospects and challenges. 1st Workshop on "Risk prognosis of environmental
SYBR Advantage qpcr Premix User Manual
SYBR Advantage qpcr Premix User Manual Cat. No. 639676 PT3883- (PR65850) Published 3 May 006 Table of Contents I. I ntroduction 4 II. List of Components 6 III. Additional Materials Required 6 IV. General
INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist
INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist Objective of PCR To provide a solution to one of the most pressing
Polymerase Chain Reaction-361 BCH
Polymerase Chain Reaction-361 BCH 1-Polymerase Chain Reaction Nucleic acid amplification is an important process in biotechnology and molecular biology and has been widely used in research, medicine, agriculture
Multiplex PCR Assay Kit Ver.2
Cat. # RR062A For Research Use Multiplex PCR Assay Kit Ver.2 Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage... 3 IV. Precautions... 3 V. Designing Primers for Multiplex
Application Notes
Application Notes Superior Performance and Flexibility In order to minimize errors in DNA replication during PCR, it is essential to choose a high-fidelity DNA polymerase enzyme. The introduction of errors
SYBR Premix Ex Taq (Tli RNaseH Plus), Bulk
Cat. # RR420L For Research Use SYBR Premix Ex Taq (Tli RNaseH Plus), Bulk Product Manual Table of Contents I. Description... 3 II. Principle... 3 III. Kit Components... 4 IV. Materials Required but not
Instructions for Use Life Science Kits & Assays
Instructions for Use Life Science Kits & Assays Product specifications 1 Product specifications The innumix qpcr SyGreen Sensitive has been developed for fast, highly reproducible real-time PCR and has
500U. Unit Definition. Storage Buffer. 10X PCR Buffer with Mg 2+
Hy-Taq 500U + dntps #EZ1012 500U Concentration: 5U/μl Contents: Hy-Taq DNA Polymerase 100μl 10xPCR Buffer(Mg 2+ Plus) 1.25ml dntps(2.5mm each) 1ml 6xLoading Buffer 1ml Store at -20 C For research only
PrimeScript RT Master Mix (Perfect Real Time)
Cat. # RR036A For Research Use PrimeScript RT Master Mix (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided... 4 IV. Storage...
Quant Reverse Transcriptase
1. Quant Reverse Transcriptase For first-strand cdna synthesis and two-step RT-PCR www.tiangen.com RT080530 Kit Contents Quant Reverse Transcriptase Contents Cat. no. ER103 ER103-02 25 rxns ER103-03 50
PrimeScript RT reagent Kit with gdna Eraser (Perfect Real Time)
For Research Use PrimeScript RT reagent Kit with gdna Eraser (Perfect Real Time) Product Manual Table of Contents I. Description... 3 II. Kit Components... 3 III. Materials Required but not Provided...
Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami
Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami hanialhadrami@kau.edu.sa www.hanialhadrami.kau.edu.sa Overview Several techniques are available to detect and analyse RNA. Examples of these techniques
Roche Molecular Biochemicals Technical Note No. 4/99
Roche Molecular Biochemicals Technical Note No. 4/99 LightCycler LightCycler-RNA Amplification Kit SYBR Green I (96 rxn) Cat. No. 2 05 37 Adaptation Protocol for Sequence-Independent Detection of RNA with
Real-time PCR Product Selection Guide
Real-time PCR Product Selection Guide Taqman Probe SYBR Green RealMOD Green AP 5x RealMOD Probe HP 5x Cat.No. 25448 / 250 rxn. / Cat.No. 25349 / 250 rxn. / RECOMMEND RealMOD Green W² 2x Real-time PCR RECOMMEND
Real Time Quantitative PCR Assay Validation, Optimization and Troubleshooting
Real Time Quantitative PCR Assay Validation, Optimization and Troubleshooting Dr Steffen Muller Field Applications Scientist Stratagene Europe Overview The Importance of Controls Validation and Optimization
SuperReal PreMix Plus (SYBR Green)
SuperReal PreMix Plus (SYBR Green) For fast, quantitative, real-time PCR using SYBR Green www.tiangen.com/en QP120627 SuperReal PreMix Plus (SYBR Green) Cat. no. FP205 Kit Contents Contents 2 SuperReal
Sensitivity vs Specificity
Viral Detection Animal Inoculation Culturing the Virus Definitive Length of time Serology Detecting antibodies to the infectious agent Detecting Viral Proteins Western Blot ELISA Detecting the Viral Genome
Recombinant DNA Technology
History of recombinant DNA technology Recombinant DNA Technology (DNA cloning) Majid Mojarrad Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists