Navigation Links
TripleMaster PCR System

e range for high fidelity PCR, optimal reaction parameters (incubation times, temperatures, concentrations of template DNA, primer, Mg2+ and enzyme mix) vary and must be determined for each primer/template pair individually.

Protocol for high fidelity PCR

Prepare two Mastermixes and keep them on ice. Mastermix 1 contains primers and template. Mastermix 2 contains enzyme, dNTPs and buffer. Combine Mastermixes 1 and 2 immediately prior to commencing cycling in order to avoid primer/template degradation by 35 exonuclease activity and non-specific primer extension at lower temperatures.
  1. Thaw the reagents listed in Table 5 carefully and keep on ice.
  2. Make sure reagents are thoroughly thawed. Mix each reagent and briefly centrifuge before use.
  3. Prepare an enzyme dilution in 1x HighFidelity Buffer as described in the appendix, if variable amounts of enzyme units will be tested.
  4. Set up Mastermix 1 in a sterile microcentrifuge tube, mix well, centrifuge and place on ice (see Table 5).
  5. Set up Mastermix 2 in a seperate sterile microcentrifuge tube, mix well, centrifuge and place on ice (see Table 5).
  6. Just before cycling, pipette Mastermix 1 and Mastermix 2 into a separate PCR tube on ice. Mix thoroughly (vortex) and centrifuge.
  7. Place immediately into a thermal cycler pre-heated at 94C and start cycling. Example of cycling profile is given for the Eppendorf Mastercycler (see Table 6).
  8. Analyze amplification products on a 0.61.5% agarose gel using appropriate DNA markers.
Table 5: Mastermix composition for high fidelity PCR
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Related biology technology :

1. TripleMaster PCR System
2. The TripleMaster PCR System for three different amplifications
3. TripleMaster PCR System: Direct Comparison with Competitors
4. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
5. TripleMaster PCR SystemThe Ideal Choice for Long-Range PCR
6. Using the TripleMaster PCR System for Robust Amplification of GC-Rich DNA Templates
7. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
8. Assess the In Vivo Activation of Signal Transduction Pathways with PathDetect Reporting Systems
9. Generate Adenovirus Vectors in E. coli by Homologous Recombination with the AdEasy Adenoviral Vector System
10. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
11. Enhanced PCR Cloning System
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: TripleMaster PCR System

(Date:8/29/2014)... 2014 Pittcon is pleased to announce ... Koichiro Matsuda, Horiba Scientific, have organized a session on ... JASIS , Asia’s largest analytical and scientific instruments ... Messe, Japan. , The four speakers in this ... Single Molecules to Single Cells” will present their use ...
(Date:8/29/2014)... Local veterinarian, Dr. Keith Clement, and his ... to participate in an investigational study of donor stem ... of this study is to determine if a single ... arthritically affected joints can help reduce pain and inflammation ... current investigational study must be older than nine months, ...
(Date:8/29/2014)... August 29, 2014 Due to a misstatement ... week, the Adult Stem Cell Technology Center, LLC ( ASCTC ... stem cell therapy research that might benefit ALS patients. ... for new stem cell technologies and therapeutic applications, FRC listed ... Bucket Challenge donors who wish not to support research ...
(Date:8/29/2014)... TX (PRWEB) August 29, 2014 ... ISO 13485:2003 certified, GAMP® 5 compliant imaging core ... an international Phase II clinical trial to assess ... , Throughout this trial, Intrinsic Imaging will provide ... limited to, protocol and charter development, site qualification, ...
Breaking Biology Technology:Fifth Annual U.S. Symposium at JASIS Organized by Pittcon’s Program Chairman Hub MacDonald 2Burnt Hills Veterinary Hospital Seeks Candidates for an Investigational Study of Stem Cells for Dogs with Arthritis 2The Adult Stem Cell Technology Center, LLC Declines ALS Ice Bucket Challenge Donors Wishing to Support Ethical Research for New Stem Cell Therapies 2The Adult Stem Cell Technology Center, LLC Declines ALS Ice Bucket Challenge Donors Wishing to Support Ethical Research for New Stem Cell Therapies 3Intrinsic Imaging Awarded Phase II Clinical Trial to Assess New Treatment for Non-Hodgkin’s Lymphoma 2
... May 17, 2011 Brian Olson didn,t exactly fit the image ... be the first to admit it. "I had just a GED ... into remedial math." With a vague notion that he ... of," Olson enrolled in basic chemistry and biology courses as a ...
... May 17, 2011 P f enex ... has submitted a Biologics Master File (BMF) to the United ... carrier protein to support the development of conjugate vaccine products ... supplying cGMP grade CRM197 carrier protein to multiple partners in ...
... Mich., May 17, 2011 Neogen Corporation (Nasdaq: ... retired senior vice president of The Kroger Co. and former ... elected to Neogen,s Board of Directors. "Since our ... quality of individuals on our Board of Directors, and the ...
Cached Biology Technology:From GED to PhD: Borough of Manhattan Community College's Brian Olson Faces Academic Deficits Only to Be Rewarded with Acceptance to CUNY's PhD Program in Biochemistry 2Pfenex Inc., Through Its Reagent Proteins Business Division, Announces Submission of Biologics Master File for Recombinant CRM197 to US FDA 2Pfenex Inc., Through Its Reagent Proteins Business Division, Announces Submission of Biologics Master File for Recombinant CRM197 to US FDA 3William T. Boehm Joins Neogen's Board of Directors 2
(Date:8/28/2014)... 28, 2014  Privacy Advocate and Senior Staff Attorney ... , joins the lineup of biometric and mobile commerce ... UnPlugged Executive Summit in Tampa, Florida ... announced speakers include Steven Rahman, Director, Technology and Strategy at ... Experian. The theme of this year,s event is Mobility ...
(Date:8/28/2014)... complex systems. Their infrastructure equipment requires ancillary sub-systems ... to supply fuel, and ICT systems for control ... is interconnected with a wider network and they ... scientists has studied various aspects of inter-network dependencies, ... in EPJ B by Gaihua Fu from Newcastle ...
(Date:8/28/2014)... kilometres of new roads will be built worldwide by ... last wildernesses, where they bring an influx of destructive ... ambitious study has created a ,global roadmap, for prioritising ... the competing demands of development and environmental protection. , ... estimates that natural importance of ecosystems and a ,road-benefits, ...
Breaking Biology News(10 mins):Biometrics UnPlugged Welcomes the Electronic Frontier Foundation's Jennifer Lynch to the Mobility at the Crossroads of Commerce and Privacy Summit 2Inter-dependent networks stress test 2Study shows where on the planet new roads should and should not go 2Study shows where on the planet new roads should and should not go 3
... to be launched on Tuesday 14 February at the Lions ... hundreds of Victorians a year by extending the time between ... the new facilities, LCDS can now store corneas at human ... Previously, corneas were held at low temperatures and had to ...
... at improving the prevention, diagnosis and treatment of cancer, ... launched this week after several years of meticulous planning. ... south Manchester area are being given the opportunity to ... year. , They are being sent invitations to take ...
... director at the Max Planck Institute of Biochemistry in ... 1990s that blocking blood vessel development in a tumour ... fundamental principle led to the development of SUTENT®, whose ... clinical use in the United States, and it is ...
Cached Biology News:Australian first to save sight 2Multi-million pound UK Biobank underway 2Multi-million pound UK Biobank underway 3Basic research leads to a novel cancer therapy 2Basic research leads to a novel cancer therapy 3Basic research leads to a novel cancer therapy 4
... CHEMICON's In Vitro Osteogenesis Assay Kit ... MC3T3-E1cells to a mature osteoblastic lineage, as ... this kit provides all the necessary reagents ... a standard plate reader. This product is ...
... of discovery races ever faster, you need a ... track. So our constant focus is on providing ... Thats why our Biomek 3000 Laboratory Automation Workstation ... as it is flexible. By integrating all aspects ...
... Cycler is the ideal instrument for budget-conscious labs ... PCR applications. Get Applied Biosystems proven ... Save lab space without sacrificing results. ... PCR Technology You Can Afford The ...
... micro 10 represents a major advance in ... the benefits of large robotic liquid handlers ... dispenser. Dispensing accurate volumes ranging from 10mL ... has unique advantages not yet found in ...
Biology Products: