Definition
RLGS is a two-dimensional gel electrophoresis method that is used as a genome scanning technique to survey genomes for copy number changes as well as changes in DNA methylation status of promoter sequences.
Characteristics
RLGS was developed as a high-speed genome scanning technique for the identification of DNA amplification as well as DNA methylation changes. For RLGS, genomic DNA is restriction digested with a landmark enzyme, usually a rare-cutting enzyme (e.g., NotI). The restriction ends are end labeled in a fill-in reaction using [α-32P]-labeled nucleotides. In the following second restriction digest (e.g., EcoRV), the DNA fragments are cut into smaller fragments suitable for separation in an agarose disk gel electrophoresis (first dimension). The next step is an in-gel restriction digestion with a third enzyme (e.g., HinfI) that generates fragments with a size range between 100 and 2,000 base pairs. DNA fragments are separated in a polyacrylamide gel (second...
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References
Costello JF, Plass C, Arap W et al (1997) Cyclin-dependent kinase 6 (CDK6) amplification in human gliomas identified using two-dimensional separation of genomic DNA. Cancer Res 57:1250–1254
Costello JF, Frühwald MC, Smiraglia DJ et al (2000) Aberrant CpG island methylation has non-random and tumor type specific patterns. Nat Genet 25:132–138
Hatada I, Hayashizaki Y, Hirotsune S et al (1991) A genomic scanning method for higher organisms using restriction sites as landmarks. Proc Natl Acad Sci U S A 88:9523–9527
Hayashizaki Y, Shibata H, Hirotsune S et al (1994) Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method. Nat Genet 6:33–40
Plass C, Shibata H, Kalcheva I et al (1996) Identification of Grf1 on mouse chromosome 9 as an imprinted gene by RLGS-M. Nat Genet 14:106–109
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© 2016 Springer-Verlag Berlin Heidelberg
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Plass, C. (2016). Restriction Landmark Genomic Scanning. In: Schwab, M. (eds) Encyclopedia of Cancer. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-46875-3_5062
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DOI: https://doi.org/10.1007/978-3-662-46875-3_5062
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