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Separation of the complementary strands of DNA fragments on polyacrylamide gels

Szalay, A. A. and Grohmann, K. and Sinsheimer, R. L. (1977) Separation of the complementary strands of DNA fragments on polyacrylamide gels. Nucleic Acids Research, 4 (5). pp. 1569-1578. ISSN 0305-1048.

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32P-labeled (in vivo) øX174 RFI DNA was restricted by Hinc II. Three aliquots of the same digest: a) nondenatured, b) heat denatured, and c) denatured by 5 mM Me-HgOH were analyzed on 3-15% acrylamide gelgradients or on 3% gels with reduced N,N'-methylene-bis-acrylamide. The autoradiography of the gels showed that the nondenatured sample migrates two times faster than the denatured samples. After denaturation each original fragment appeared as a doublet. Using in vitro synthesized RFI DNA labeled only in negative strand with 32P we could identify the position of the negative strand in each denatured doublet. The single strand DNA fragments could be recovered from the gel slices on a semi-preparative scale by electrophoresis into dialysis tubing.

Item Type:Article
Additional Information:Copyright © 1977 Oxford University Press. Received January 5, 1977. This work was supported by U.S. Public Health Service Grant No. GM 13554. We thank Professors N. Davidson and S. K. Dube for their critical discussions.
Record Number:CaltechAUTHORS:SZAnar77
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4015
Deposited By: Tony Diaz
Deposited On:24 Jul 2006
Last Modified:26 Dec 2012 08:57

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