Encyclopedia of Cancer

Living Edition
| Editors: Manfred Schwab

Restriction Landmark Genomic Scanning

Living reference work entry
DOI: https://doi.org/10.1007/978-3-642-27841-9_5062-2

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...

Keywords

Methylation Status Imprint Gene NotI Site Restriction Landmark Genomic Scanning Intensity Fragment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. 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 57:1250–1254PubMedGoogle Scholar
  2. Costello JF, Frühwald MC, Smiraglia DJ et al (2000) Aberrant CpG island methylation has non-random and tumor type specific patterns. Nat Genet 25:132–138CrossRefGoogle Scholar
  3. 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 88:9523–9527CrossRefPubMedPubMedCentralGoogle Scholar
  4. 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 6:33–40CrossRefPubMedGoogle Scholar
  5. 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 14:106–109CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.German Cancer Research Center (DKFZ)HeidelbergGermany