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Ultrasensitive amplicon barcoding for next-generation sequencing facilitating sequence error and amplification-bias correction.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Ahmed, Ibrahim 
Tucci, Felicia A 
Aflalo, Aure 
Smith, Kenneth GC 
Bashford-Rogers, Rachael JM 

Abstract

The ability to accurately characterize DNA variant proportions using PCR amplification is key to many genetic studies, including studying tumor heterogeneity, 16S microbiome, viral and immune receptor sequencing. We develop a novel generalizable ultrasensitive amplicon barcoding approach that significantly reduces the inflation/deflation of DNA variant proportions due to PCR amplification biases and sequencing errors. This method was applied to immune receptor sequencing, where it significantly improves the quality and estimation of diversity of the resulting library.

Description

Keywords

Bias, DNA, DNA Barcoding, Taxonomic, DNA Primers, Gene Library, High-Throughput Nucleotide Sequencing, Humans, Nucleic Acid Amplification Techniques, Polymerase Chain Reaction, Sequence Analysis, DNA

Journal Title

Sci Rep

Conference Name

Journal ISSN

2045-2322
2045-2322

Volume Title

10

Publisher

Springer Science and Business Media LLC
Sponsorship
Wellcome Trust (106068/Z/14/Z)