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

Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963-5. doi: 10.1073/pnas.80.13.3963.

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.

MeSH terms

  • Autoradiography
  • Base Sequence*
  • Buffers
  • DNA*
  • Deoxyadenine Nucleotides
  • Electrophoresis, Polyacrylamide Gel / methods
  • Sulfur Radioisotopes
  • Thionucleotides

Substances

  • Buffers
  • Deoxyadenine Nucleotides
  • Sulfur Radioisotopes
  • Thionucleotides
  • 2'-deoxyadenosine 5'-O-(1-thiotriphosphate)
  • DNA