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:8/21/2014)... MD (PRWEB) August 21, 2014 DNA ... making progress in its development of JRC-LAMP-vax, an innovative ... cedar pollen. Phase IC studies began this month in ... to hay fever. , The Japanese red cedar releases ... people in Japan. Mountain Cedar pollen wreaks similar havoc ...
(Date:8/21/2014)... , August 21, 2014 ... http://www.researchandmarkets.com/research/3zh3q8/global_hormone ) has announced the addition of ...  report to their offering.       ... as hormone therapy, is a means of ... produced by the body. This type of ...
(Date:8/21/2014)... , Aug. 21, 2014  Decision Resources Group ... Brazil and Mexico ... of several newly approved agents, the anticipated label ... and the launch of emerging biologics will substantially ... particular, the emerging IL-5 inhibitors will introduce a ...
(Date:8/21/2014)... , Aug. 21, 2014  Four-dimensional (4-D) printing ... based on external stimuli such as changes in ... a disruptive effect in multiple industries.  In the ... landscape by printing objects ranging from human organs ... sectors. Logo - http://photos.prnewswire.com/prnh/20140820/138289 ...
Breaking Biology Technology:Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 2Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 3Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 4Global Hormone Replacement Therapy Market 2014-2018: Key Vendors are Actavis, Bayer, Merck & Co, Novartis and Novo Nordisk 2Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 2Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 3Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 2Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 3Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 4
... VisEn Medical Inc., a leader in fluorescence in vivo ... of its new Cat-K FAST (TM) ... with disease progression and therapeutic response in vivo . ... therapeutic target in a range of bone-related diseases, including cancer metastasis ...
... 7 Mylan Inc. (Nasdaq: MYL ) today ... from the U.S. Food and Drug Administration (FDA) for its ... , , Bicalutamide Tablets are the generic version of ... of approximately $322 million for the 12 months ending March ...
... MATEO, Calif., July 7 King Pharmaceuticals, Inc. (NYSE: ... today announced that on July 2, 2009, King met with the ... Letter regarding the New Drug Application (NDA) for REMOXY(R). The ... to resubmit the REMOXY(R) NDA and to address all FDA comments ...
Cached Biology Technology:VisEn Launches New Cat-K FAST(TM) Fluorescence Molecular Imaging Agent 2King and Pain Therapeutics Announce REMOXY(R) NDA Update 2King and Pain Therapeutics Announce REMOXY(R) NDA Update 3
(Date:8/21/2014)... unknown process for harvesting energy and producing oxygen from ... University scientist has discovered. The discovery lays the foundation ... energy from the Sun, and understanding dense blooms like ... worldwide. A paper describing the discovery will be published ... journal Science on 21 August 2014. , ...
(Date:8/21/2014)... aimed at improving the food intake, health and quality ... is among three projects at the University of Waterloo ... Canadian Institutes of Health Research (CIHR). , Professor ... at Waterloo, and a Schlegel research chair in nutrition ... investigate why many Canadians living in long-term care homes ...
(Date:8/21/2014)... Ever heard of the water window? It consists of ... are not absorbed by the water in biological tissues. ... develop coherent radiations within the water window. These could ... a high-contrast image of the biological samples or to ... study identifies the physical mechanism needed to efficiently generate ...
Breaking Biology News(10 mins):Hot-spring bacteria reveal ability to use far-red light for photosynthesis 2Hot-spring bacteria reveal ability to use far-red light for photosynthesis 3Grants will fund landmark aging research at Waterloo 2Water window imaging opportunity 2
... to be satisfied? Less than half as much as ... snacking study. Using chocolate chips, apple pie, and potato ... Wansink designed a study to determine if people who ... hungrier or satisfied fifteen minutes after eating., ,Two groups ...
... in the American Chemical Society,s (ACS,) award-winning Global ... to improve the sensitivity of the test often used to ... possessions of passengers at airport security screening stations. Based ... ACS, The Journal of Physical Chemistry C , the ...
... Aviv Prof. Nathan Nelson of Tel Aviv University,s ... Center has been awarded the 2012 Israel Prize in ... awarded in Jerusalem on the eve of Israel,s Independence ... and molecular biology, Prof. Nelson is internationally renowned for ...
Cached Biology News:New American Chemical Society podcast: Boosting the sensitivity of airport security screening 2
Practical tool for easy, effective sealing of 0.2ml PCR tubes and plates using cap strips....
... by BioLegend, the Quansys Biosciences Q-Plex Array ... of cytokines/chemokines for human, mouse, and rat ... a fully quantitative ELISA-based test where 4 ... each well of a 96-well plate in ...
The strip cap tool is a used for properly seating 8- and 12-cap strips in microplates or strip tubes. A grooved channel on one side is designed to seat domed caps, while the flat edge on the opposite...
... The expertise that made the original Gene Pulser ... system available has been applied in the Gene ... The modular design of the Gene Pulser II ... accessory that has the widest range of settings ...
Biology Products: