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Long-Range Polymerase Chain Reaction

  • William Waggott
Protocol
  • 134 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Conventional polymerase chain reaction (PCR) enables reliable amplification of 3–4 kb of DNA (1) while attempts at optimization has enabled 15.6 kb of λ DNA to be amplified (2). The maximum amplifiable length of PCR is limited by the low fidelity of the Thermus aquaticus (Taq) DNA polymerase (3), the most commonly used thermostable polymerase. It is believed that inadvertent nucleotide misincorporations during the PCR extension steps cause chain terminations (3). The Taq polymerase lacks proofreading properties (4) and thus is unable to correct such misincorporations. The higher extension KM value for a misincorporated nucleotide is thought to cause detachment of the Taq polymerase from template DNA.

Keywords

Polymerase Chain Reaction Polymerase Chain Reaction Primer Anaplastic Large Cell Lymphoma Myotonic Dystrophy Exonuclease Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • William Waggott
    • 1
  1. 1.Department of Cellular Sciences, John Radcliffe HospitalUniversity of OxfordUK

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