Mutation assay using single-molecule real-time (SMRT™) sequencing technology

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抄録

Introduction: We present here a simple, phenotype-independent mutation assay using a PacBio RSII DNA sequencer employing single-molecule real-time (SMRT) sequencing technology. Salmonella typhimurium YG7108 was treated with the alkylating agent N-ethyl-N-nitrosourea (ENU) and grown though several generations to fix the induced mutations, the DNA was extracted and the mutations were analyzed by using the SMRT DNA sequencer. Results: The ENU-induced base-substitution frequency was 15.4 per Megabase pair, which is highly consistent with our previous results based on colony isolation and next-generation sequencing. The induced mutation spectrum (95% G:C→A:T, 5% A:T→G:C) is also consistent with the known ENU signature. The base-substitution frequency of the control was calculated to be less than 0.12 per Megabase pair. A current limitation of the approach is the high frequency of artifactual insertion and deletion mutations it detects. Conclusions: Ultra-low frequency base-substitution mutations can be detected directly by using the SMRT DNA sequencer, and this technology provides a phenotype-independent mutation assay.

収録刊行物

  • Genes and Environment

    Genes and Environment (37), 2015-09-01

    BioMed Central Ltd.

各種コード

  • NII論文ID(NAID)
    120005749959
  • 本文言語コード
    ENG
  • 資料種別
    journal article
  • ISSN
    1880-7062
  • データ提供元
    IR 
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