Quantitation of in situ hybridization of ribosomal ribonucleic acids to human diploid cells. Journal Article uri icon

Overview

abstract

  • The hybridization of 5S and 28S ribosomal RNAs to human fibroblast and leukocyte cells was used as a model system to quantitative the technique of in situ hybridization for human diploid cell types. Quantitation consisted of counting (scoring) the number of grains formed over both interphase nuclei and metaphase chromosomes on slides after various hybridization procedures. The average number of grains/nucleus per slide was then used to determine hybridization percentages. As with nitrocellulose filter hybridizations the kinetics of in situ hybridizations can be fit with a single first-order rate-constant. However, the in situ hybridization rate was approximately 10 times slower than the corresponding filter hybridization rate. The efficiency of in situ hybridization was found to range between 5 and 15% for both leukocyte and fibroblast cell types and for both metaphase and interphase nuclei. Determination of the parameters of the in situ hybridization reaction of ribosomal RNAs to diploid chromosomes define the experimental conditions needed for the localization of single copy genes to diploid chromosomes.

publication date

  • January 1, 1980

Date in CU Experts

  • March 13, 2015 12:57 PM

Full Author List

  • Coté BD; Uhlenbeck OC; Steffensen DM

author count

  • 3

citation count

  • 0

Other Profiles

International Standard Serial Number (ISSN)

  • 0009-5915

Electronic International Standard Serial Number (EISSN)

  • 1432-0886

Additional Document Info

start page

  • 349

end page

  • 367

volume

  • 80

issue

  • 3