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
prolekare.web.journal.doi_sk:
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:
DNA – Biology and life sciences – Genetics – Genomics – Genome analysis – Biochemistry – Nucleic acids – Computational biology – Research and analysis methods – Molecular biology – Gene identification and analysis – Molecular biology techniques – DNA construction – Molecular probe techniques – Artificial gene amplification and extension – Polymerase chain reaction – Specimen preparation and treatment – DNA electrophoresis – Electrophoretic techniques – Sequencing techniques – Transcriptome analysis – Mechanical treatment of specimens – Sonication – DNA sequencing – Next-generation sequencing – Mutation detection – DNA libraries – DNA manipulations – DNA fragment ligation – Probe hybridization – DNA hybridization – Ultrasonication
Zdroje
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