Labeling of the centromeric region on human chromosome 8 by in situ hybridization

Heinz Ulrich G Weier, Joe Gray, Hans Dieter Kleine

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Probe DNA that binds preferentially to the centromeric region of human chromosomes 8 was synthesized. Alpha satellite probe DNA molecules were selectively amplified from sorter-purified human chromosomes 8 by in vitro DNA amplification using the polymerase chain reaction (PCR). Probe labeling was performed during PCR by incorporation of biotinylated deoxyuridine. In situ hybridization of unpurified probe DNA comprised of alpha satellite monomer and higher molecular weight DNA fragments with metaphase chromosome spreads showed binding to the centromeric regions of numerous chromosomes. However, blocking with unlabeled total human alphoid DNA dramatically reduced crosshybridization to chromosomes other than 8. Under these conditions, the degenerate probe DNA allowed unambiguous visualization of domains occupied by centromeric DNA of chromosome 8 in metaphase spreads and interphase cell nuclei, thus greatly facilitating the detection of numerical chromosome aberrations in tumor cells. In situ hybridization of size-fractionated alpha satellite DNA identified the monomeric fraction as the major cause of crosshybridization. Alpha satellite dimers and higher molecular weight DNA fragments showed relatively high specificity for human chromosomes 8.

Original languageEnglish (US)
Pages (from-to)489-494
Number of pages6
JournalHuman Genetics
Volume87
Issue number4
DOIs
StatePublished - Aug 1991
Externally publishedYes

Fingerprint

Chromosomes, Human, Pair 8
Human Chromosomes
In Situ Hybridization
DNA Probes
DNA
Satellite DNA
Metaphase
Chromosomes
Molecular Weight
Polymerase Chain Reaction
Deoxyuridine
Interphase
Cell Nucleus
Chromosome Aberrations
Neoplasms

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

Labeling of the centromeric region on human chromosome 8 by in situ hybridization. / Weier, Heinz Ulrich G; Gray, Joe; Kleine, Hans Dieter.

In: Human Genetics, Vol. 87, No. 4, 08.1991, p. 489-494.

Research output: Contribution to journalArticle

Weier, Heinz Ulrich G ; Gray, Joe ; Kleine, Hans Dieter. / Labeling of the centromeric region on human chromosome 8 by in situ hybridization. In: Human Genetics. 1991 ; Vol. 87, No. 4. pp. 489-494.
@article{0aaefc71b6e143aeb29a2bbab8dede39,
title = "Labeling of the centromeric region on human chromosome 8 by in situ hybridization",
abstract = "Probe DNA that binds preferentially to the centromeric region of human chromosomes 8 was synthesized. Alpha satellite probe DNA molecules were selectively amplified from sorter-purified human chromosomes 8 by in vitro DNA amplification using the polymerase chain reaction (PCR). Probe labeling was performed during PCR by incorporation of biotinylated deoxyuridine. In situ hybridization of unpurified probe DNA comprised of alpha satellite monomer and higher molecular weight DNA fragments with metaphase chromosome spreads showed binding to the centromeric regions of numerous chromosomes. However, blocking with unlabeled total human alphoid DNA dramatically reduced crosshybridization to chromosomes other than 8. Under these conditions, the degenerate probe DNA allowed unambiguous visualization of domains occupied by centromeric DNA of chromosome 8 in metaphase spreads and interphase cell nuclei, thus greatly facilitating the detection of numerical chromosome aberrations in tumor cells. In situ hybridization of size-fractionated alpha satellite DNA identified the monomeric fraction as the major cause of crosshybridization. Alpha satellite dimers and higher molecular weight DNA fragments showed relatively high specificity for human chromosomes 8.",
author = "Weier, {Heinz Ulrich G} and Joe Gray and Kleine, {Hans Dieter}",
year = "1991",
month = "8",
doi = "10.1007/BF00197174",
language = "English (US)",
volume = "87",
pages = "489--494",
journal = "Human Genetics",
issn = "0340-6717",
publisher = "Springer Verlag",
number = "4",

}

TY - JOUR

T1 - Labeling of the centromeric region on human chromosome 8 by in situ hybridization

AU - Weier, Heinz Ulrich G

AU - Gray, Joe

AU - Kleine, Hans Dieter

PY - 1991/8

Y1 - 1991/8

N2 - Probe DNA that binds preferentially to the centromeric region of human chromosomes 8 was synthesized. Alpha satellite probe DNA molecules were selectively amplified from sorter-purified human chromosomes 8 by in vitro DNA amplification using the polymerase chain reaction (PCR). Probe labeling was performed during PCR by incorporation of biotinylated deoxyuridine. In situ hybridization of unpurified probe DNA comprised of alpha satellite monomer and higher molecular weight DNA fragments with metaphase chromosome spreads showed binding to the centromeric regions of numerous chromosomes. However, blocking with unlabeled total human alphoid DNA dramatically reduced crosshybridization to chromosomes other than 8. Under these conditions, the degenerate probe DNA allowed unambiguous visualization of domains occupied by centromeric DNA of chromosome 8 in metaphase spreads and interphase cell nuclei, thus greatly facilitating the detection of numerical chromosome aberrations in tumor cells. In situ hybridization of size-fractionated alpha satellite DNA identified the monomeric fraction as the major cause of crosshybridization. Alpha satellite dimers and higher molecular weight DNA fragments showed relatively high specificity for human chromosomes 8.

AB - Probe DNA that binds preferentially to the centromeric region of human chromosomes 8 was synthesized. Alpha satellite probe DNA molecules were selectively amplified from sorter-purified human chromosomes 8 by in vitro DNA amplification using the polymerase chain reaction (PCR). Probe labeling was performed during PCR by incorporation of biotinylated deoxyuridine. In situ hybridization of unpurified probe DNA comprised of alpha satellite monomer and higher molecular weight DNA fragments with metaphase chromosome spreads showed binding to the centromeric regions of numerous chromosomes. However, blocking with unlabeled total human alphoid DNA dramatically reduced crosshybridization to chromosomes other than 8. Under these conditions, the degenerate probe DNA allowed unambiguous visualization of domains occupied by centromeric DNA of chromosome 8 in metaphase spreads and interphase cell nuclei, thus greatly facilitating the detection of numerical chromosome aberrations in tumor cells. In situ hybridization of size-fractionated alpha satellite DNA identified the monomeric fraction as the major cause of crosshybridization. Alpha satellite dimers and higher molecular weight DNA fragments showed relatively high specificity for human chromosomes 8.

UR - http://www.scopus.com/inward/record.url?scp=0025834759&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025834759&partnerID=8YFLogxK

U2 - 10.1007/BF00197174

DO - 10.1007/BF00197174

M3 - Article

VL - 87

SP - 489

EP - 494

JO - Human Genetics

JF - Human Genetics

SN - 0340-6717

IS - 4

ER -