High-fidelity PCR (CAT#: STEM-MB-0204-WXH)

Introduction

High-fidelity PCR is a modifies PCR method that utilizes a DNA polymerase with a low error rate and results in a high degree of accuracy in the replication of the DNA of interest.<br />Such enzymes have a significant binding affinity for the correct nucleoside triphosphate during amplification.<br />In the case of an incorrect binding in the polymerase active site, incorporation is slowed due to the architecture of the active site complex.<br />Highfidelity amplification is essential for experiments whose outcome depends upon the correct DNA sequence like cloning, SNP analysis, NGS applications.

Add to Cart Please Inquire

Principle Applications Procedure Materials Notes Related Products

Principle

Replicative polymerases achieve high fidelity of DNA replication by employing several mechanisms: (1) sensing proper geometry of correct base pair, (2) slowing down catalysis in case of a mismatch, and (3) partitioning the mismatched primer to exonuclease active site.

Applications

For some applications such as cloning and site directed mutagenesis, or when preparing a template for sequencing, the number of mutations introduced during PCR needs to be kept to a minimum.

Procedure

General procedure:
1. Denaturation
2. Annealing
3. Extension

Other recommended products

Analysis of LINC00538 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-0425-LGZ)
Analysis of KIT D816V Mutation by RT-qPCR (CAT#: STEM-MT-0006-LGZ)
Analysis of OR52E8 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-2372-LGZ)
Analysis of Bovine Hepervirus 2 (BHV-2) by Real-Time PCR Method (CAT#: STEM-MB-2763-LGZ)
Analysis of C1orf226 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-0247-LGZ)
Analysis of LIPF Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-2087-LGZ)
Detection of Unknown β-globin Gene Mutations by RT-qPCR (CAT#: STEM-MT-0037-LGZ)
Analysis of Sendai Virus (SV) by Real-Time PCR Method (CAT#: STEM-MB-4055-LGZ)