Reducing chimera formation during PCR amplification to ensure accurate genotyping

R.P. Smyth; T.E. Schlub; A. Grimm; V. Venturi; A. Chopra; S. Mallal; M.P. Davenport; J. Mak

doi:10.1016/j.gene.2010.08.009

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Reducing chimera formation during PCR amplification to ensure accurate genotyping

Journal article

Peer reviewed

Reducing chimera formation during PCR amplification to ensure accurate genotyping

R.P. Smyth, T.E. Schlub, A. Grimm, V. Venturi, A. Chopra, S. Mallal, M.P. Davenport and J. Mak

Gene, Vol.469(1-2), pp.45-51

2010

DOI: https://doi.org/10.1016/j.gene.2010.08.009

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Abstract

Measurements of population diversity are fundamental to the reconstruction of the evolutionary and epidemiological history of organisms. Commonly used protocols to measure population diversity using the polymerase chain reaction (PCR) are prone to the introduction of artificial chimeras. These are often difficult to detect and can confound the correct interpretation of results due to the false generation of recombinants when the underlying DNA sample contains multiple distinct templates. This study presents a standardised procedure to suppress the formation of artificial chimeras during PCR amplification. The solution is based on the accurate determination of the efficiency and end point of the log-linear phase of a PCR. This procedure will facilitate the generation of data sets that more accurately reflect the underlying population diversity rather than artifacts introduced by the process itself.

Details

Title: Reducing chimera formation during PCR amplification to ensure accurate genotyping
Authors/Creators: R.P. Smyth (Author/Creator) - Burnet Institute
T.E. Schlub (Author/Creator) - UNSW Sydney
A. Grimm (Author/Creator) - UNSW Sydney
V. Venturi (Author/Creator) - UNSW Sydney
A. Chopra (Author/Creator) - Royal Perth Hospital
S. Mallal (Author/Creator) - Royal Perth Hospital
M.P. Davenport (Author/Creator) - UNSW Sydney
J. Mak (Author/Creator) - Burnet Institute
Publication Details: Gene, Vol.469(1-2), pp.45-51
Publisher: Elsevier BV
Identifiers: 991005544841307891
Copyright: © 2010 Elsevier B.V.
Murdoch Affiliation: Centre for Clinical Immunology and Biomedical Statistics
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration
Citation topics: 3 Agriculture, Environment & Ecology; 3.83 Bioengineering; 3.83.167 Microbial Diversity
Web Of Science research areas: Genetics & Heredity
ESI research areas: Molecular Biology & Genetics