Analysis of genomic aberrations using comparative genomic hybridization of metaphase chromosomes.

Melanie Carless

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Original languageEnglish
Pages (from-to)177-202
Number of pages26
JournalMethods in molecular biology (Clifton, N.J.)
Volume523
StatePublished - 2009
Externally publishedYes

Fingerprint

Comparative Genomic Hybridization
Metaphase
Chromosomes
DNA
Fluorescence
Polymerase Chain Reaction
Oncogenes
Fluorescence Microscopy
Paraffin
Formaldehyde
In Situ Hybridization
Blood Cells
Genome
Cell Line
Neoplasms

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Analysis of genomic aberrations using comparative genomic hybridization of metaphase chromosomes. / Carless, Melanie.

In: Methods in molecular biology (Clifton, N.J.), Vol. 523, 2009, p. 177-202.

Research output: Contribution to journalArticle

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abstract = "Comparative genomic hybridization (CGH) allows the global screening of copy number aberrations within a sample. Specifically, large (>20 mb) deletions and amplifications are detected, which are likely to indicate regions harboring tumor suppressor and oncogenes. CGH involves the extraction of test and reference (karyotypically normal) DNA. These samples are whole-genome amplified by DOP-PCR and then differentially labeled with fluorophores via nick translation. Test and reference samples are competitively hybridized to normal metaphase chromosomes. The relative amount of each DNA that binds to a chromosomal locus is indicative of the abundance of that DNA. Thus, if a chromosomal region is amplified, the test DNA will outcompete the reference DNA for binding and fluorescence will indicate amplification. Conversely, if a region is deleted, more reference DNA will bind and fluorescence will indicate a deletion. The following chapter outlines the protocols used for CGH analysis of metaphase chromosomes. These protocols include metaphase chromosome slide preparation, DNA extraction (from blood, cell lines and microdissected formalin-fixed paraffin-embedded tissue), DOP-PCR, nick translation, in situ hybridization and fluorescence microscopy and image analysis.",
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