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Feasibility of mapping low-multiplicity genes by in situ hybridization. [/sup 125/I and tritium tracers]

Journal Article:

Abstract

The assignment of human hemoglobin loci to 2q and Bq was followed by objections based on the low specific activity of their /sup 3/H-mRNA, estimated as somewhere between 100 and 1000 dpm/..mu..g. With that preparation, the time required for one disintegration per molecule is between 80,000 and 8000 years. The consideration that globin loci may contain as many as 10 copies does not help. In view of these incontestably valid objections, it is instructive to compare the foregoing experiment with another in which the chromosomes were hybridized with cDNA copied from human reticulocyte mRNA by means of the reverse transcriptase from avian myeloblastosis virus. Our preparation had a specific activity of 1.4 x 10/sup 8/ dpm/..mu..g, requiring less than a month per disintegration per molecule. Despite the rather more favorable anticipated exposure time, successful localization was by no means expected. The use of cDNA precluded the enzymatic removal of background; the concentration of DNA applied to the slides, 0.03 ..mu..g/ml, was unfavorable for hybridization; and the copying reaction, primed with oligo-dT, probably attached stretches up to T/sub 20/ to the structural sequence, with the possibility that these might hybridize to uninteresting chromosomal regions. Results indicate that the original assignments of  More>>
Publication Date:
Jan 01, 1975
Product Type:
Journal Article
Reference Number:
EDB-78-039652
Resource Relation:
Journal Name: Cytogenet. Cell Genet.; (Switzerland); Journal Volume: 14
Subject:
59 BASIC BIOLOGICAL SCIENCES; HEMOGLOBIN; HYBRIDIZATION; BIOCHEMICAL REACTION KINETICS; MAN; GENETIC MAPPING; BIOLOGICAL EFFECTS; CHROMOSOMES; DNA; IN VITRO; MESSENGER-RNA; MOLECULAR BIOLOGY; RIBOSOMES; TRACER TECHNIQUES; VIRUSES; ANIMALS; CARBOXYLIC ACIDS; CELL CONSTITUENTS; GLOBIN; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; ISOTOPE APPLICATIONS; KINETICS; MAMMALS; MICROORGANISMS; NUCLEIC ACIDS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANOIDS; PARASITES; PIGMENTS; PORPHYRINS; PRIMATES; PROTEINS; REACTION KINETICS; RNA; VERTEBRATES; 550401* - Genetics- Tracer Techniques
OSTI ID:
5340802
Research Organizations:
Columbia Univ., New York
Country of Origin:
Switzerland
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: CGCGB
Submitting Site:
TIC
Size:
Pages: 279-281
Announcement Date:

Journal Article:

Citation Formats

Atwood, K C, Henderson, A S, Kacian, D, and Eicher, E M. Feasibility of mapping low-multiplicity genes by in situ hybridization. [/sup 125/I and tritium tracers]. Switzerland: N. p., 1975. Web.
Atwood, K C, Henderson, A S, Kacian, D, & Eicher, E M. Feasibility of mapping low-multiplicity genes by in situ hybridization. [/sup 125/I and tritium tracers]. Switzerland.
Atwood, K C, Henderson, A S, Kacian, D, and Eicher, E M. 1975. "Feasibility of mapping low-multiplicity genes by in situ hybridization. [/sup 125/I and tritium tracers]." Switzerland.
@misc{etde_5340802,
title = {Feasibility of mapping low-multiplicity genes by in situ hybridization. [/sup 125/I and tritium tracers]}
author = {Atwood, K C, Henderson, A S, Kacian, D, and Eicher, E M}
abstractNote = {The assignment of human hemoglobin loci to 2q and Bq was followed by objections based on the low specific activity of their /sup 3/H-mRNA, estimated as somewhere between 100 and 1000 dpm/..mu..g. With that preparation, the time required for one disintegration per molecule is between 80,000 and 8000 years. The consideration that globin loci may contain as many as 10 copies does not help. In view of these incontestably valid objections, it is instructive to compare the foregoing experiment with another in which the chromosomes were hybridized with cDNA copied from human reticulocyte mRNA by means of the reverse transcriptase from avian myeloblastosis virus. Our preparation had a specific activity of 1.4 x 10/sup 8/ dpm/..mu..g, requiring less than a month per disintegration per molecule. Despite the rather more favorable anticipated exposure time, successful localization was by no means expected. The use of cDNA precluded the enzymatic removal of background; the concentration of DNA applied to the slides, 0.03 ..mu..g/ml, was unfavorable for hybridization; and the copying reaction, primed with oligo-dT, probably attached stretches up to T/sub 20/ to the structural sequence, with the possibility that these might hybridize to uninteresting chromosomal regions. Results indicate that the original assignments of the human globin loci are correct.}
journal = {Cytogenet. Cell Genet.; (Switzerland)}
volume = {14}
journal type = {AC}
place = {Switzerland}
year = {1975}
month = {Jan}
}