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In-gel multiple displacement amplification of long DNA fragments diluted to the single molecule level
https://repo.qst.go.jp/records/47663
https://repo.qst.go.jp/records/47663b7144c35-2823-4f25-9fa9-4505ea17981a
Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2017-03-15 | |||||
タイトル | ||||||
タイトル | In-gel multiple displacement amplification of long DNA fragments diluted to the single molecule level | |||||
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 | journal 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 |
Michikawa, Yuichi
× Michikawa, Yuichi× Sugahara, Keisuke× Suga, Tomo× Ohtsuka, Yoshimi× Ishikawa, Kenichi× Ishikawa, Atsuko× Shiomi, Naoko× Shiomi, Tadahiro× Iwakawa, Mayumi× Imai, Takashi× 道川 祐市× 菅原 圭亮× 菅 智× 荘司 好美× 石川 顕一× 石川 敦子× 塩見 尚子× 塩見 忠博× 岩川 眞由美× 今井 高志 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | The isolation and multiple genotyping of long individual DNA fragments are needed to obtain haplotype information for diploid organisms. Limiting dilution of sample DNA followed by multiple displacement amplification is a useful technique but is restricted to short (<5 kb) DNA fragments. In the current study, a novel modification was applied to overcome these problems. A limited amount of cellular DNA was carefully released from intact cells into a mildly heated alkaline agarose solution and mixed thoroughly. The solution was then gently aliquoted and allowed to solidify while maintaining the integrity of the diluted DNA. Exogenously provided Phi29 DNA polymerase was used to perform consistent genomic amplification with random hexameric oligonucleotides within the agarose gels. Simple heat melting of the gel allowed recovery of the amplified materials in a solution of the polymerase chain reaction (PCR)-ready form. The haplotypes of seven SNPs spanning 240 kb of the DNA surrounding the human ATM gene region on chromosome 11 were determined for 10 individuals, demonstrating the feasibility of this new method. | |||||
書誌情報 |
Analytical Biochemistry 巻 383, 号 2, p. 151-158, 発行日 2008-08 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 0003-2697 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1016/j.ab.2008.08.011 |