Rapid denaturation improves chromosome morphology and permits multiple hybridizations during fluorescence in situ hybridization

K. H. Ramesh, M. J. Macera, R. S. Verma

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Denaturation of chromosomal DNA for fluorescence in situ hybridization (FISH) is an essential step in a procedure associated with a number of variables. In our experience, shorter denaturation time in 70% formamide/2 x SSC at 72 C provides sufficient denaturation, where the hydrogen bonds are broken between the purines and pyrimidines of the double helix. This shortened exposure improves retention of morphology of human chromosomes from lymphocytes, aminocytes, fibroblasts and bone marrow, and allows the same metaphases to be denatured repeatedly and rehybridized with different probes. This approach is useful in investigations where sample volume is limited.

Original languageEnglish (US)
Pages (from-to)141-143
Number of pages3
JournalBiotechnic and Histochemistry
Volume72
Issue number3
DOIs
Publication statusPublished - May 1997
Externally publishedYes

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Keywords

  • Chromosomes
  • Denaturation
  • FISH

ASJC Scopus subject areas

  • Histology
  • Medical Laboratory Technology

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