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 Citations (Scopus)

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
StatePublished - May 1997
Externally publishedYes

Fingerprint

Nucleic Acid Denaturation
Pyrimidines
Purines
Human Chromosomes
Metaphase
Fluorescence In Situ Hybridization
Hydrogen
Fibroblasts
Chromosomes
Bone Marrow
Lymphocytes
formamide

Keywords

  • Chromosomes
  • Denaturation
  • FISH

ASJC Scopus subject areas

  • Anatomy
  • Applied Microbiology and Biotechnology

Cite this

Rapid denaturation improves chromosome morphology and permits multiple hybridizations during fluorescence in situ hybridization. / Ramesh, K.h.; Macera, M. J.; Verma, R. S.

In: Biotechnic and Histochemistry, Vol. 72, No. 3, 05.1997, p. 141-143.

Research output: Contribution to journalArticle

@article{fe2fd630357240a5a92bfda3f25a6696,
title = "Rapid denaturation improves chromosome morphology and permits multiple hybridizations during fluorescence in situ hybridization",
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.",
keywords = "Chromosomes, Denaturation, FISH",
author = "K.h. Ramesh and Macera, {M. J.} and Verma, {R. S.}",
year = "1997",
month = "5",
language = "English (US)",
volume = "72",
pages = "141--143",
journal = "Biotechnic and Histochemistry",
issn = "1052-0295",
publisher = "Informa Healthcare",
number = "3",

}

TY - JOUR

T1 - Rapid denaturation improves chromosome morphology and permits multiple hybridizations during fluorescence in situ hybridization

AU - Ramesh, K.h.

AU - Macera, M. J.

AU - Verma, R. S.

PY - 1997/5

Y1 - 1997/5

N2 - 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.

AB - 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.

KW - Chromosomes

KW - Denaturation

KW - FISH

UR - http://www.scopus.com/inward/record.url?scp=0030951888&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030951888&partnerID=8YFLogxK

M3 - Article

C2 - 9187736

AN - SCOPUS:0030951888

VL - 72

SP - 141

EP - 143

JO - Biotechnic and Histochemistry

JF - Biotechnic and Histochemistry

SN - 1052-0295

IS - 3

ER -