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Nucleic Acids Research, 1989, Vol. 17, No. 6 2181-2186
© 1989

CHEMISTRY

Non-radioactive chemical sequencing of biotin labelled DNA

Peter Richterich

Fakultät fü;r Biologic, Universität Konstanz D-7750 Konstanz, FRG

Received January 17, 1989. Revised January 23, 1989. Accepted January 23, 1989.

Methods for the nonradioactive chemical sequencing of DNA are described. A biotin marker molecule, attached chemically to an oligonucleotide primer or enzymatically in an endfilling reaction of restriction enzyme sites, is stable during the base-specific chemical modification and strand scission reactions. Following fragment separation by direct blotting electrophoresis, the membrane bound sequence pattern can be visualized by a streptavidin-bridged enzymatic color reaction. The biotin labeling is also applicable for DNA sequencing by random degradation of phosphothioates, thus showing to be a universal label for nonradioactive DNA sequencing.


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