Long fragment polymerase chain reaction

Eng Wee Chua, Simran Maggo, Martin A. Kennedy

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Polymerase chain reaction (PCR) is an oft-used preparatory technique in amplifying specific DNA regions for downstream analysis. The size of an amplicon was initially limited by errors in nucleotide polymerization and template deterioration during thermal cycling. A variant of PCR, designated long-range PCR, was devised to counter these drawbacks and enable the amplification of large fragments exceeding a few kb. In this chapter we describe a protocol for long-range PCR, which we have adopted to obtain products of 6.6, 7.2, 13, and 20 kb from human genomic DNA samples.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages65-74
Number of pages10
Volume1620
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1620
ISSN (Print)1064-3745

Fingerprint

Polymerase Chain Reaction
DNA
Polymerization
Nucleotides
Hot Temperature

Keywords

  • Agarose gel electrophoresis
  • Long amplicons
  • Long-range polymerase chain reaction (PCR)
  • Proofreading enzyme
  • Taq DNA polymerase
  • Thermal cycling

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Chua, E. W., Maggo, S., & Kennedy, M. A. (2017). Long fragment polymerase chain reaction. In Methods in Molecular Biology (Vol. 1620, pp. 65-74). (Methods in Molecular Biology; Vol. 1620). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7060-5_3

Long fragment polymerase chain reaction. / Chua, Eng Wee; Maggo, Simran; Kennedy, Martin A.

Methods in Molecular Biology. Vol. 1620 Humana Press Inc., 2017. p. 65-74 (Methods in Molecular Biology; Vol. 1620).

Research output: Chapter in Book/Report/Conference proceedingChapter

Chua, EW, Maggo, S & Kennedy, MA 2017, Long fragment polymerase chain reaction. in Methods in Molecular Biology. vol. 1620, Methods in Molecular Biology, vol. 1620, Humana Press Inc., pp. 65-74. https://doi.org/10.1007/978-1-4939-7060-5_3
Chua EW, Maggo S, Kennedy MA. Long fragment polymerase chain reaction. In Methods in Molecular Biology. Vol. 1620. Humana Press Inc. 2017. p. 65-74. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7060-5_3
Chua, Eng Wee ; Maggo, Simran ; Kennedy, Martin A. / Long fragment polymerase chain reaction. Methods in Molecular Biology. Vol. 1620 Humana Press Inc., 2017. pp. 65-74 (Methods in Molecular Biology).
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