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Synthesis, Cleavage and Sequence Analysis of DNA Complementary to the 26 S Messenger RNA of Sindbis Virus

Rice, Charles M. and Strauss, James H. (1981) Synthesis, Cleavage and Sequence Analysis of DNA Complementary to the 26 S Messenger RNA of Sindbis Virus. Journal of Molecular Biology, 150 (3). pp. 315-340. ISSN 0022-2836.

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Conditions for synthesis of long complementary DNA transcripts of Sindbis virus 26 S and 49 S RNA in high yield have been developed. This single-stranded complementary DNA could be cut with HaeIII, HhaI, RsaI or TaqI to give reproducible patterns of discrete, virus-specific fragments that were suitable for subsequent end-labeling and direct sequence analysis. Using these methods we present the strategy used for obtaining nearly the entire 26 S RNA sequence from complementary DNA synthesized in vitro. This approach should prove useful for sequence analysis of any purified RNA available in microgram quantities.

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Additional Information:© 1981 Academic Press Inc. (London) Ltd. Received 7 March 1981. We would like to thank E. Lenches for preparing the chick fibroblasts; T. E. Crowley for help in the early stages of this work; H. V. Huang, T. Hunkapiller, M. Douglas and R. Douglas for writing computer programs; J. R. Bell for the preparation of some of the Figures; and L. Hood and T. Maniatis for their generosity in lending us supplies and equipment. This work was supported by grants PCM 80-22830 from the National Science Foundation, by grants GM06965 and AI10793 from the National Institutes of Health, and by Biomedical Research Support grant 507RR 07003 from the National Institutes of Health. One author (C.M.R.) was supported by training grant GM00086 from the National Institutes of Health.
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NSFPCM 80-22830
NIHGM 06965
NIHAI 10793
NIH507RR 07003
NIH Predoctoral FellowshipGM-00086
Issue or Number:3
Record Number:CaltechAUTHORS:20160516-151804415
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Official Citation:Charles M. Rice, James H. Strauss, Synthesis, cleavage and sequence analysis of DNA complementary to the 26 S messenger RNA of Sindbis virus, Journal of Molecular Biology, Volume 150, Issue 3, 1981, Pages 315-340, ISSN 0022-2836, (
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:67141
Deposited By: Donna Wrublewski
Deposited On:23 May 2016 22:33
Last Modified:03 Oct 2019 10:03

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