JaLCDOI 10.18926/AMO/30857
フルテキストURL fulltext.pdf
著者 Ikeda, Shogo| Yamamoto, Mihoko| Nagao, Kazutaka| Zhang, Bo| Watanabe, Sekiko| Oda, Takuzo|
抄録 <p>Non-radioactive hybridization probes were prepared using the M13 phage vector and the universal sequencing primer. The probe sequence to be used was first cloned into the M13 vector, and the minus strand of the template DNA was then synthesized with the Klenow fragment of E. coli DNA polymerase I in the presence of the biotinylated nucleotide, biotin-11-dUTP, as a label. Resultant DNA was heavily biotinylated, and made up of the entire minus strand of the template DNA. The long tag sequence derived from the M13 vector may increase the sensitivity of the detection. The biotinylated hybrids were visualized with the streptavidin-alkaline phosphatase conjugate and chromogenic substrates. As shown by Southern hybridization, the probe prepared in this way could be used to detect less than 1 pg of target sequence and a single copy gene sequence in human genomic DNA within several hours of signal development.</p>
キーワード non-rradioctive probe biotin nucleotide M13 phage DNA universal sequencing primer Southern hybridization
Amo Type Article
発行日 1989-08
出版物タイトル Acta Medica Okayama
43巻
4号
出版者 Okayama University Medical School
開始ページ 197
終了ページ 202
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 2678902
Web of Sience KeyUT A1989AP79100001