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

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...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 80; no. 13; pp. 3963 - 3965
Main Authors Biggin, M D, Gibson, T J, Hong, G F
Format Journal Article
LanguageEnglish
Published United States National Acad Sciences 01.07.1983
Subjects
Autoradiography
Base Sequence
Buffers
Deoxyadenine Nucleotides
DNA
Electrophoresis, Polyacrylamide Gel - methods
Sulfur Radioisotopes
Thionucleotides
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Summary: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.
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ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.80.13.3963