Synthesis of RNA from cellulose-bound complementary DNA
Radioactively labeled RNAs were synthesized from cellulose-bound cDNA templates using Escherichia coli RNA polymerase. Hybridization of this RNA to excess unlabeled cDNA approached 100%, indicating the complementarity of product and template. The average length of the RNA product, as determined by f...
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Published in | The Journal of biological chemistry Vol. 252; no. 16; pp. 5615 - 5618 |
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Main Authors | , |
Format | Journal Article |
Language | English |
Published |
United States
American Society for Biochemistry and Molecular Biology
25.08.1977
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Subjects | |
Online Access | Get full text |
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Summary: | Radioactively labeled RNAs were synthesized from cellulose-bound cDNA templates using Escherichia coli RNA polymerase. Hybridization
of this RNA to excess unlabeled cDNA approached 100%, indicating the complementarity of product and template. The average
length of the RNA product, as determined by formamide gels, was approximately 40% of the template length. Hybridization of
unlabeled globin RNA produced by this technique to labeled globin cDNA indicated the population of RNA sequences represented
at least 80% of the template sequences. Approximately 30% of the RNA product by mass contains poly(A) tails as determined
by binding to oligo(dT)-cellulose. The template can be reused for several cycles of synthesis with little loss of synthetic
capability and therefore, can amplify the amount of mRNA initially used to produce the template. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0021-9258 1083-351X |
DOI: | 10.1016/S0021-9258(17)40066-4 |