Genome Scanning Method; Restriction Landmark Genomic Scanning (RLGS)

Abstract

The restriction landmark genomic scanning (RLGS) technology is a method for high-speed survey of the presence or absence of restriction landmarks throughout a genome and the measurement of their copy number based on the new concept that restriction enzyme sites are used as landmarks. RLGS employs (1) direct end-labeling of the genomic DNA digested with a restriction enzyme and (2) high-resolution, two-dimensional electrophoresis. RLGS has the following advantages. (1) High speed–scanning ability. Thousands of restriction landmarks can be scanned simultaneously. (2) Scanning field can be extended by the use of different kinds of landmarks in an additional series of electrophoresis; (3) This method can be applied to any organism because direct labeling of restriction enzyme sites, and not the hybridization procedure, is employed as a detection system. (4) Spot intensity reflects the copy number of the restriction landmark. Thus, haploid and diploid genomic DNAs can be distinguished. (5) Using a methylation-sensitive enzyme, the methylated state of genomic DNA can be screened. Thus, RLGS is a very useful system not only for genome mapping but also for various studies in biological and medical fields, such as research for cancer development, aging, and so on. Keywords: Genome Scanning; Restriction Landmark; Blocking (in RLGS)