Navigation Links
Pfu DNA Polymerase as a Hot Start Alternative

Traditional hot start additives unnecessary


Kirk B. Nielson Janice Cline Holly Hogrefe
Stratagene Cloning Systems, Inc.


This article describes the important, novel application of Stratagenes Pfu DNA polymerase for minimizing PCR cold start mismatched primer-extension reactions. These mismatched extension reactions result in the synthesis of undesirable artifactual PCR products. The problem of mismatched extension reactions may be alleviated by using Taq DNA polymerase in conjunction with hot start techniques, which prevent nucleotide incorporation prior to the first PCR denaturation step. Pfu DNA polymerase,, * which exhibits minimal activity at problematic lower temperatures, obviates the need for Taq DNA polymerase-based hot start procedures.

Pfu DNA polymerase has the lowest error rate of any thermostable DNA polymerase studied to date.1,2 This feature makes it the preferred polymerase for techniques that require high-fidelity DNA synthesis by the polymerase chain reaction (PCR). Applications for Pfu DNA polymerase include gene cloning and expression, sequencing and site-directed mutagenesis. Pfu DNA polymerase exhibits exceptionally high thermostability3 and requires a relatively high temperature for optimal activity (Topt75C), characteristics that have significant implications in PCR applications. Most notable is Pfu DNA polymerases relatively low activity, in comparison to other commonly used PCR enzymes, at temperatures ranging from 22 to 50C. Minimal polymerase activity at these temperatures should result in fewer mispaired primer-extension reactions than occur with Taq DNA polymerase, which exhibits significantly higher activity at temperatures used in the primer-annealing phase of PCR (~50C).

The relatively high thermostability and Topt <
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9

Related biology technology :

1. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
2. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
3. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
4. Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications
5. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
6. High-Fidelity PCR with a Novel Polymerase Mixture
7. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
8. Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance
9. The Use of MasterTaq Polymerase for PCR with Humic Material-Contaminated DNA
10. HotStarTaq DNA Polymerase
11. C. therm. Polymerase One-Step RT-PCR System
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Pfu DNA Polymerase Hot Start Alternative

(Date:8/29/2014)... /PRNewswire/ - iCo Therapeutics ("iCo" or "the Company") ... for the six months ended June 30, 2014. ... dollars and presented under International Financial Reporting Standards ... , Announced top-line results related to the ... subjects enrolled in the Phase 2 iDEAL Study ...
(Date:8/29/2014)... (PRWEB) August 29, 2014 Intrinsic ... 13485:2003 certified, GAMP® 5 compliant imaging core lab, ... international Phase II clinical trial to assess a ... Throughout this trial, Intrinsic Imaging will provide comprehensive ... to, protocol and charter development, site qualification, site ...
(Date:8/28/2014)... Ind. A new research platform uses ... of tiny structures undergoing stress and heating, ... improve designs for microelectronics and batteries., This ... information about how heating and the surface ... properties. Researchers have discussed the merits of ...
(Date:8/28/2014)... , Aug. 28, 2014   SunTrust Robinson Humphrey ... industry veterans to its equity research team in ... ongoing expansion in equity research demonstrates our commitment ... insights to enhance their investment decision making," said ... STRH.  "We continue to make significant investments in ...
Breaking Biology Technology:iCo Therapeutics Announces Second Quarter 2014 Financial Results 2iCo Therapeutics Announces Second Quarter 2014 Financial Results 3iCo Therapeutics Announces Second Quarter 2014 Financial Results 4Intrinsic Imaging Awarded Phase II Clinical Trial to Assess New Treatment for Non-Hodgkin’s Lymphoma 2New analytical technology reveals 'nanomechanical' surface traits 2New analytical technology reveals 'nanomechanical' surface traits 3SunTrust Robinson Humphrey Expands Equity Research in Healthcare and Energy 2SunTrust Robinson Humphrey Expands Equity Research in Healthcare and Energy 3
... , , , , , Multiporator / Electroporator 2510 , , , ... Transformation Protocol , Protocol No. 4308 915.501 04/2002 ... , , , , , ... , Cell type , ...
... Kits , , ... Eppendorf Perfectprep Plasmid DNA Isolation Kits are designed to generate , ... automated fluorescent , sequencing, transfection, PCR, and other extremely ... In order to compare yields between our kits and the leading competitor, ...
... , Other important parameters for optimizing electroporation., , , , , , ... , Pulse length , ... Adjustment , of the electroporation buffer , ... Influence , of temperature , ...
Cached Biology Technology:Actinomyces viscosus 2Eppendorf Perfectprep Plasmid DNA Isolation , Kits 2Eppendorf Perfectprep Plasmid DNA Isolation , Kits 3Optimizing electroporation parameters 2
(Date:8/28/2014)... the developmental on-off switch for Streptomyces , a ... of the world,s naturally derived antibiotic medicines. , ... is possible to manipulate this switch to make nature,s ... August 28 in Cell , found that a ... a larger protein called BldD ultimately controls whether a ...
(Date:8/28/2014)... FASEB MARC (Maximizing Access to Research Careers) ... the American College of Sports Medicine,s Conference on ... in Miami, Florida. These awards are meant ... and scientists from underrepresented groups into the mainstream ... the participation of young scientists at the American ...
(Date:8/28/2014)... Dyslexia, the most commonly diagnosed learning disability in ... that occurs when the regions of the brain ... The use of non-invasive functional neuroimaging tools has ... dyslexia. However, most prior work has focused on ... a gap in our understanding of how multiple ...
Breaking Biology News(10 mins):Small molecule acts as on-off switch for nature's antibiotic factory 2Small molecule acts as on-off switch for nature's antibiotic factory 3Dyslexic readers have disrupted network connections in the brain 2
... National data indicate that milk consumption has declined ... nutritional quality has more than doubled. Although this suggests ... in children,s diets, a new study suggests that in ... related to changes in their consumption of sodas and ...
... at the California Institute of Technology (Caltech) pioneered ... the immune system and neurodevelopmental disorders such as ... postmortem brains and of individuals with autism, as ... between alterations in the immune system and autism ...
... is a vital nutrient for pig growth, but pigs ... the University of Illinois has determined how adding various ... how pigs digest the phosphorus in four different feed ... bottom lines as well as for the environment. ...
Cached Biology News:Sugar-sweetened drinks are not replacing milk in kid's diets 2Sugar-sweetened drinks are not replacing milk in kid's diets 3Caltech researchers find evidence of link between immune irregularities and autism 2Caltech researchers find evidence of link between immune irregularities and autism 3Helping pigs to digest phosphorus 2Helping pigs to digest phosphorus 3
Mouse monoclonal antibody raised against a partial recombinant ALAS2. NCBI Entrez Gene ID = ALAS2...
Mouse monoclonal antibody raised against a partial recombinant QPCT. NCBI Entrez Gene ID = QPCT...
Mouse monoclonal antibody raised against a partial recombinant STAU2. NCBI Entrez Gene ID = STAU2...
... gun system, 220-240 V, is used for ... This handheld device employs an adjustable helium ... biomaterial-coated gold microcarriers from the inner wall ... target cells. This system has a 2 ...
Biology Products: