Restriction landmark genomic scanning

被引:12
作者
Ando, Yoshinari
Hayashizaki, Yoshihide
机构
[1] RIKEN, Yokohama Inst, Frontier Res Syst, Funct RNA Res Program,Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan
[2] RIKEN, Yokohama Inst, Genom Sci Ctr, Genome Explorat Res Grp, Yokohama, Kanagawa 2300045, Japan
关键词
D O I
10.1038/nprot.2006.350
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Restriction landmark genomic scanning (RLGS) is a method to detect large numbers of restriction landmarks in a single experiment. It is based on the concept that restriction enzyme sites can serve as landmarks throughout a genome. RLGS uses direct end-labeling of the genomic DNA digested with a rare-cutting restriction enzyme and high-resolution two-dimensional electrophoresis. Compared with the conventional gene-detection technologies, such as Southern blot analysis and PCR, RLGS has the following advantages even though it needs specially designed instruments: high-efficiency scanning capacity, scanning extensibility by using alternate restriction enzyme combinations, applicability to any organism, a spot intensity that reflects the copy number of restriction landmarks, and the ability, by using a methylation-sensitive enzyme, to screen the methylated state of genomic DNA. The RLGS protocol can be accomplished in 5 days to 2 weeks.
引用
收藏
页码:2774 / 2783
页数:10
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