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High-resolution melting curve analysis for rapid detection of mutations in a Medaka TILLING library by Tomoko Ishikawa; Yasuhiro Kamei; Shinji Otozai; Jinhyong Kim; Ayuko Sato; Yoshikazu Kuwahara; Minoru Tanaka; Tomonori Deguchi; Hidenori Inohara; Tohru Tsujimura; Takeshi Todo is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is High-resolution melting curve analysis for rapid detection of mutations in a Medaka TILLING library about?

## Abstract ## Background During the last two decades, DNA sequencing has led to the identification of numerous genes in key species; however, in most cases, their functions are still unknown. In this situation, reverse genetics is the most suitable method to assign function to a gene. TILLING (Targeting Induced Local Lesions IN Genomes) is a reverse-genetic strategy that combines random chemical mutagenesis with high-throughput discovery of the induced mutations in target genes. The method has been applied to a variety of plant and animal species. Screening of the induced mutations is the most important step in TILLING. Currently, direct sequencing or nuclease-mediated screening of heteroduplexes is widely used for detection of mutations in TILLING. Both methods are useful, but the costs are substantial and turnaround times are relatively long. Thus, there is a need for an alternative method that is of higher throughput and more cost effective. ## Results In this study, we developed a high resolution melting (HRM) assay and evaluated its effectiveness for screening ENU-induced mutations in a medaka TILLING library. We had previously screened mutations in the__p53__gene by direct s

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Author

Tomoko Ishikawa; Yasuhiro Kamei; Shinji Otozai; Jinhyong Kim; Ayuko Sato; Yoshikazu Kuwahara; Minoru Tanaka; Tomonori Deguchi; Hidenori Inohara; Tohru Tsujimura; Takeshi Todo

Publisher

BioMed Central; Springer (Biomed Central Ltd.); London: BioMed Central, [2000]-; Springer Science and Business Media LLC (ISSN 1471-2199)

Published

2010

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)