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Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination.

Proceedings of the National Academy of Sciences of the United States of America (1983-07-01)
M D Biggin, T J Gibson, G F Hong
ABSTRACT

Two methods for increasing the length of DNA sequence data that can be read off a polyacrylamide gel are described. We have developed a rapid way to pour a buffer concentration gradient gel that, by altering the vertical band separation on an autoradiograph, allows more sequence to be obtained from a gel. We also show that the use of deoxyadenosine 5'-(alpha-[35S]thio)triphosphate as the label incorporated in dideoxynucleotide sequence reactions increases the sharpness of the bands on an autoradiograph and so increases the resolution achieved.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tris-Borate-EDTA buffer, 5× concentrate, powder blend

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