Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2967:181-192.
doi: 10.1007/978-1-0716-3358-8_15.

Long-Range Polymerase Chain Reaction

Ping Siu Kee  1 , Harsheni Karunanathie  2 , Simran D S Maggo  1   3 , Martin A Kennedy  1 , Eng Wee Chua  4
Affiliations

Long-Range Polymerase Chain Reaction

Ping Siu Kee et al. Methods Mol Biol. 2023.

Abstract

Polymerase chain reaction (PCR) is a laboratory technique used to amplify a targeted region of DNA, demarcated by a set of oligonucleotide primers. Long-range PCR is a form of PCR optimized to facilitate the amplification of large fragments. Using the adapted long-range PCR protocol described in this chapter, we were able to generate PCR products of 6.6, 7.2, 13, and 20 kb from human genomic DNA samples. For some of the long PCRs, successful amplification was not possible without the use of PCR enhancers. Thus, we also evaluated the impact of some enhancers on long-range PCR and included the findings as part of this updated chapter.

Keywords: Agarose gel electrophoresis; DNA polymerase; Long amplicons; Long-range polymerase chain reaction (PCR); PCR additives; PCR enhancers; Pharmacogenetics; Primer design; Proofreading enzyme; Thermal cycling.

PubMed Disclaimer

References

    1. Mullis KB, Faloona FA (1987) Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. In: Methods in enzymology, vol 155. Academic Press, pp 335–350
    1. Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239(4839):487–491 - DOI - PubMed
    1. Collins F, Galas D (1993) A new five-year plan for the US human genome project. Science 262(5130):43–46 - DOI - PubMed
    1. Delidow BC, Lynch JP, Peluso JJ, White BA (1993) Polymerase chain reaction. In: PCR Protocols. Humana Press, Totowa, NJ, pp 1–29
    1. Long S (2022). Digital PCR: methods and applications in infectious diseases. Methods (San Diego, Calif.)

LinkOut - more resources