Screening for Alagille syndrome mutations in the JAG1 and NOTCH2 genes using denaturing high-performance liquid chromatography

Hazuki Samejima, Chiharu Torii, Rika Kosaki, Kenji Kurosawa, Hiroshi Yoshihashi, Koji Muroya, Nobuhiko Okamoto, Yoriko Watanabe, Tomoki Kosho, Michiru Kubota, Osamu Matsuda, Miwa Goto, Kosuke Izumi, Takao Takahashi, Kenjiro Kosaki

研究成果: Article査読

6 被引用数 (Scopus)

抄録

Mutations in the JAG1 gene and the NOTCH2 gene cause Alagille syndrome. At present, however, genetic testing of Alagille syndrome is not commonly applied in clinical settings because the currently available assays are technically and financially demanding, mainly because of the size of the genes. In the present study, we optimized the highly sensitive and specific mutation scanning method automated denaturing high-performance liquid chromatography (DHPLC) to analyze the entire coding region of JAG1 and NOTCH2. The coding region was amplified by 69 primer pairs, all of which have the same cycling conditions, aliquoted on a 96-well format PCR plate. In this manner, all the exons were simultaneously amplified using a single block in a thermal cycler. We then wrote a computer script to analyze each segment of JAG1 and NOTCH2 by DHPLC in a serial manner using conditions that were optimized for each amplicon. The implementation of this screening method for JAG1 and NOTCH2 will help medical geneticists confirm their clinical impressions and provide accurate genetic counseling to the patients with Alagille syndrome and their families.

本文言語English
ページ(範囲)216-227
ページ数12
ジャーナルGenetic Testing
11
3
DOI
出版ステータスPublished - 2007 9月 1

ASJC Scopus subject areas

  • 遺伝学(臨床)

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