A method for counting PCR template molecules with application to next-generation sequencing.

A method for counting PCR template molecules with application to next-generation sequencing.
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DOI:
10.1093/nar/gkr217
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发表时间:
2011-07
影响因子:
14.9
通讯作者:
Lichtenstein CP
Lichtenstein CP
中科院分区:
生物学2区
文献类型:
--
作者:
Casbon JA;Osborne RJ;Brenner S;Lichtenstein CP

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Amplification by polymerase chain reaction is often used in the preparation of template DNA molecules for next-generation sequencing. Amplification increases the number of available molecules for sequencing but changes the representation of the template molecules in the amplified product and introduces random errors. Such changes in representation hinder applications requiring accurate quantification of template molecules, such as allele calling or estimation of microbial diversity. We present a simple method to count the number of template molecules using degenerate bases and show that it improves genotyping accuracy and removes noise from PCR amplification. This method can be easily added to existing DNA library preparation techniques and can improve the accuracy of variant calling.
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