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High-frequency, low-coverage "false positives" mutations may be true in GS Junior sequencing studies.

ABSTRACT: The GS Junior sequencer provides simplified procedures for library preparation and data processing. Errors in pyrosequencing generate some biases during library construction and emulsion PCR amplification. False-positive mutations are identified by related characteristics described in the manufacturer's manual, and some detected mutations may have 'borderline' characteristics when they are detected in few reads or at low frequency. Among these mutations, however, some may be true positives. This study aimed to improve the accuracy of identifying true positives among mutations with borderline false-positive characteristics detected with GS Junior sequencing. Mutations with the borderline features were tested for validity with Sanger sequencing. We examined 10 mutations detected in coverages <20-fold at frequencies >30% (group A) and 16 mutations detected in coverages >20-fold at frequencies?

SUBMITTER: Yang Z

PROVIDER: S-EPMC5653793 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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High-frequency, low-coverage "false positives" mutations may be true in GS Junior sequencing studies.

Yang Zhiliang Z Sun Guilian G

Scientific reports 20171023 1

The GS Junior sequencer provides simplified procedures for library preparation and data processing. Errors in pyrosequencing generate some biases during library construction and emulsion PCR amplification. False-positive mutations are identified by related characteristics described in the manufacturer's manual, and some detected mutations may have 'borderline' characteristics when they are detected in few reads or at low frequency. Among these mutations, however, some may be true positives. This ...[more]

PMID: 29062110

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