PMID: 6159588Sep 25, 1980Paper

Sequence complexity of cDNA transcribed from a diverse mRNA population

Nucleic Acids Research
J Van Ness, W E Hahn

Abstract

Mouse liver poly(A)+mRNA was reverse transcribed using oligo-p(dT) or random oligonucleotides as primers to yield cDNA about equal to the mass of the template RNA. The size profile of the oligo-p(dT)-primedd cDNA was similar to that of the template RNA. RNA or cDNA driven saturation annealing of labeled single copy genomic DNA (scDNA) showed that 2% of the scDNA was complementary in either case indicating the sequence complexity of cDNA was equivalent to that of the template mRNA. These results establish for the first time that cDNA represents essentially all of the sequence complexity of a diverse template RNA population in which individual mRNA species are present in vastly different concentrations. RNA driven hydridization of the cDNA showed that about 40% of the cDNA mass represents most of the sequence complexity of the template RNA. Also, kinetics of this hybridization indicate a complexity of 58,000 kb for the template RNA, a value similar to that obtained by scDNA hybridization. We conclude that appropriately characterized cDNA probes can be used to make valid qualitative and quantitative comparisons of the complex, infrequent class mRNAs of different cells and tissues.

References

Oct 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·D L Kacian, J C Myers
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Citations

Jul 15, 1981·Developmental Biology·L S Levy, J E Manning
Feb 1, 1990·Genetic Analysis, Techniques and Applications·J WelshA Efstratiadis
Aug 25, 1983·Nucleic Acids Research·R J Milner, J G Sutcliffe
Mar 1, 1989·The Plant Cell·N C NielsenR B Goldberg
Nov 18, 2008·Applied and Environmental Microbiology·Roberto A RodríguezCharles P Gerba
Jun 1, 1982·Cell·R L Fischer, R B Goldberg
May 27, 1983·Science·N Chaudhari, W E Hahn

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