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

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• 作者：Tomonari Matsuda ; Shun Matsuda ; Masami Yamada
• 关键词：PacBio RSII DNA sequencer ; Single ; molecule real ; time (SMRT) sequencing technology ; Mutation assay
• 刊名：Genes and Environment
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：37
• 期：1
• 全文大小：1039KB
• 参考文献：1.Matsuda T, Takamune M, Matsuda Y, Yamada M. A pilot study for the mutation assay using a high-throughput DNA sequencer. Genes and Environ. 2013;35:53鈥?.CrossRef
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  3.Kennedy SR, Schmitt MW, Fox EJ, Kohrn BF, Salk JJ, Ahn EH, et al. Detecting ultralow-frequency mutations by duplex sequencing. Nat Protoc. 2014;9:2586鈥?06.PubMed Central PubMed CrossRef
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  5.Matsuda T. Anticipated mutation assay using single-molecule real-time (SMRT TM) sequencing technology. Genes and Environ. 2010;32:21鈥?.CrossRef
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  8.Yamada M, Matsui K, Sofuni T, Nohmi T. New tester strains of Salmonella typhimurium lacking O6-methylguanine DNA methyltransferases and highly sensitive to mutagenic alkylating agents. Mutat Res. 1997;381:15鈥?4.PubMed CrossRef
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  10.Richardson KK, Richardson FC, Crosby RM, Swenberg JA, Skopek TR. DNA base changes and alkylation following in vivo exposure of Escherichia coli to N-methyl-N-nitrosourea or N-ethyl-N-nitrosourea. Proc Natl Acad Sci U S A. 1987;84:344鈥?.PubMed Central PubMed CrossRef
  
• 作者单位：Tomonari Matsuda (1) (3)
  Shun Matsuda (1)
  Masami Yamada (2)
  
  1. Research Center for Environmental Quality Management, Kyoto University, Shiga, Japan
  3. Tomonari Matsuda, Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga, 520-0811, Japan
  2. National institute of Health Sciences, Tokyo, Japan
  
• 刊物类别：Human Genetics;
• 刊物主题：Human Genetics;
• 出版者：BioMed Central
• ISSN：1880-7062

文摘

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.