PCR-free whole exome sequencing: Cost-effective and efficient in detecting rare mutations
Autoři:
Izumi Yamaguchi aff001; Takashi Watanabe aff001; Osamu Ohara aff001; Yoshinori Hasegawa aff001
Působiště autorů:
Laboratory of Clinical Omics Research, Department of Applied Genomics, Kazusa DNA Research Institute, Chiba, Japan
aff001
Vyšlo v časopise:
PLoS ONE 14(9)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0222562
Souhrn
In this study, we describe the development of a PCR-free whole exome sequencing method. Using this method, 2 μg DNA was sufficient for library preparation for whole exome sequencing. Furthermore, the method is simple and makes use of a commercial kit, with additional step of concentrating the captured library by ethanol precipitation. The accuracy of the PCR-free method was found to be equivalent to that of unique molecular identifier-corrected analysis method, which is the commonly used method to detect rare mutations. Thus, the PCR-free whole exome sequencing method is cost-effective as well as efficient in detecting rare mutations.
Klíčová slova:
Biology and life sciences – Genetics – DNA – DNA libraries – DNA electrophoresis – Gene identification and analysis – Mutation detection – Genomics – Genome analysis – Transcriptome analysis – Biochemistry – Nucleic acids – Molecular biology – Molecular biology techniques – Artificial gene amplification and extension – Polymerase chain reaction – DNA construction – DNA manipulations – DNA fragment ligation – Molecular probe techniques – Probe hybridization – Computational biology – Research and analysis methods – DNA hybridization – Sequencing techniques – DNA sequencing – Next-generation sequencing – Specimen preparation and treatment – Mechanical treatment of specimens – Sonication – Ultrasonication – Electrophoretic techniques
Zdroje
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Článek vyšel v časopise
PLOS One
2019 Číslo 9
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