Nonspecific PCR amplification by high-fidelity polymerases: implications for next-generation sequencing of AFLP markers.

Details

Serval ID
serval:BIB_B174F94BFB88
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Nonspecific PCR amplification by high-fidelity polymerases: implications for next-generation sequencing of AFLP markers.
Journal
Molecular Ecology Resources
Author(s)
Brelsford A., Collin H., Perrin N., Fumagalli L.
ISSN
1755-0998 (Electronic)
ISSN-L
1755-098X
Publication state
Published
Issued date
2012
Peer-reviewed
Oui
Volume
12
Number
1
Pages
123-127
Language
english
Abstract
High-fidelity 'proofreading' polymerases are often used in library construction for next-generation sequencing projects, in an effort to minimize errors in the resulting sequence data. The increased template fidelity of these polymerases can come at the cost of reduced template specificity, and library preparation methods based on the AFLP technique may be particularly susceptible. Here, we compare AFLP profiles generated with standard Taq and two versions of a high-fidelity polymerase. We find that Taq produces fewer and brighter peaks than high-fidelity polymerase, suggesting that Taq performs better at selectively amplifying templates that exactly match the primer sequences. Because the higher accuracy of proofreading polymerases remains important for sequencing applications, we suggest that it may be more effective to use alternative library preparation methods.
Keywords
Amplified Fragment Length Polymorphism Analysis, Animals, Cyprinidae/genetics, DNA-Directed DNA Polymerase/metabolism, Gene Library, Oryza sativa/genetics, Polymerase Chain Reaction/instrumentation, Polymerase Chain Reaction/methods, Taq Polymerase/metabolism
Pubmed
Web of science
Create date
18/10/2011 11:21
Last modification date
20/08/2019 16:20
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