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 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.
D001345:Autoradiography; D001483:Base Sequence; D002021:Buffers; D004247:DNA; D003838:Deoxyadenine Nucleotides; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D013462:Sulfur Radioisotopes; D013873:Thionucleotides
D002021:Buffers; D003838:Deoxyadenine Nucleotides; D013462:Sulfur Radioisotopes; D013873:Thionucleotides; C015109:2'-deoxyadenosine 5'-O-(1-thiotriphosphate); D004247:DNA