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

Détails

ID Serval
serval:BIB_B174F94BFB88
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Nonspecific PCR amplification by high-fidelity polymerases: implications for next-generation sequencing of AFLP markers.
Périodique
Molecular Ecology Resources
Auteur⸱e⸱s
Brelsford A., Collin H., Perrin N., Fumagalli L.
ISSN
1755-0998 (Electronic)
ISSN-L
1755-098X
Statut éditorial
Publié
Date de publication
2012
Peer-reviewed
Oui
Volume
12
Numéro
1
Pages
123-127
Langue
anglais
Résumé
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.
Mots-clé
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
Création de la notice
18/10/2011 11:21
Dernière modification de la notice
20/08/2019 16:20
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