Amplification efficiency of thermostable DNA polymerases

The amplification efficiencies of several polymerase chain reaction (PCR) enzymes were compared using real-time quantitative PCR with SYBR Green I detection. Amplification data collected during the exponential phase of PCR are highly reproducible, and PCR enzyme performance comparisons based upon ef...

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Bibliographic Details
Published in:Analytical biochemistry 2003-10, Vol.321 (2), p.226-235
Main Authors: Arezi, Bahram, Xing, Weimei, Sorge, Joseph A, Hogrefe, Holly H
Format: Article
Language:English
Subjects:
Base Sequence
DNA-Directed DNA Polymerase - chemistry
DNA-Directed DNA Polymerase - metabolism
Enzyme Stability
GC Rich Sequence
Hot Temperature
Molecular Sequence Data
Organic Chemicals - chemistry
PCR enzymes
Pfu DNA polymerase
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - standards
Quality Control
Real-time PCR
Taq DNA polymerase
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Summary:The amplification efficiencies of several polymerase chain reaction (PCR) enzymes were compared using real-time quantitative PCR with SYBR Green I detection. Amplification data collected during the exponential phase of PCR are highly reproducible, and PCR enzyme performance comparisons based upon efficiency measurements are considerably more accurate than those based on endpoint analysis. DNA polymerase efficiencies were determined under identical conditions using five different amplicon templates that varied in length or percentage GC content. Pfu- and Taq-based formulations showed similar efficiencies when amplifying shorter targets (
ISSN:0003-2697
1096-0309
DOI:10.1016/S0003-2697(03)00465-2