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Consideration of conditions for extremely deep sequencing to detect mutations in plant DNA
https://repo.qst.go.jp/records/85291
https://repo.qst.go.jp/records/85291eae813ae-f3fb-4afb-8af0-289e08bfc045
| Item type | 一般雑誌記事 / Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2022-03-15 | |||||
| タイトル | ||||||
| タイトル | Consideration of conditions for extremely deep sequencing to detect mutations in plant DNA | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
| 資源タイプ | article | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Yutaka, Ono
× Yutaka, Ono× Satoshi, Kitamura× Katsuya, Sato× Yutaka, Ono× Satoshi, Kitamura× Katsuya, Sato |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | To establish an easy and quick method for evaluating frequency of mutations occurred in plant DNA, we have tried to apply maximum-depth sequencing (MDS), an extremely deep sequencing method that enables to detect extremely rare mutations through next generation sequencing (NGS) with an error correction. The procedure of the MDS was previously described. Briefly, each molecule of a target DNA extracted from plants was labelled with a distinct DNA barcode, lineally amplified using a single primer and DNA polymerase, and subjected to NGS. Because original mutations that existed in plant DNA will be conserved among NGS reads harboring identical barcodes, they can be distinguished from erroneous mutations caused by PCR or NGS. In this work, we have prepared 4 test reactions to test 1) dual targeting in one reaction, 2) effect of pre-treatment of DNA repair enzymes, and 3) sensitivity of this method by inputting low amount of DNA with a known mutation. | |||||
| 書誌情報 |
QST Takasaki Annual Report 2020 p. 77, 発行日 2022-03 |
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