Journal
NUCLEIC ACIDS RESEARCH
Volume 39, Issue 12, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkr217
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Funding
- Wellcome Trust [II-3A-0809-10009]
- Population Genetics Technologies Ltd.
- National Institute for Health Research [II-3A-0809-10009] Funding Source: researchfish
- National Institutes of Health Research (NIHR) [II-3A-0809-10009] Funding Source: National Institutes of Health Research (NIHR)
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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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