Restriction Landmark Genomic and cDNA Scanning

Restriction landmark genomic scanning (RLGS) is a method for the high-speed survey of the presence or absence of restriction landmarks throughout a genome. RLGS also provides for the measurement of the copy number of restriction landmarks based on the concept that restriction enzyme sites can serve as landmarks. RLGS employs direct end-labeling of the genomic DNA digested with a restriction enzyme and high-resolution, two-dimensional electrophoresis. Compared with the conventional technologies for detecting the genes, such as Southern blot analysis and polymerase chain reaction (PCR), RLGS has the following advantages even though it needs specially designed instruments: (1) high-speed scanning ability – thousands of restriction landmarks can be scanned simultaneously; (2) applicability to any organism – RLGS employs direct labeling of restriction enzyme sites, not a hybridization procedure; (3) a spot intensity that reflects the copy number of the restriction landmarks; and (4) the ability, by using a methylation-sensitive enzyme, to screen the methylated state of genomic DNA. RLGS is a very useful system not only for genome mapping, but also in a variety of biological and medical fields, such as cancer research and the growth and development of various organisms throughout all stages of the life cycle. RLGS can also be applied for the visualization of mRNAs [restriction landmark cDNA scanning (RLCS)]. In one procedure, RLCS can scan the expression of several hundred mRNAs.

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