Chemical DNA sequencing in the opposite direction to the dideoxy chain termination method, using a single preparation of M13 single-stranded DNA.
A strategy has been developed allowing the use of a single preparation of single-stranded DNA clones for chemical DNA sequencing in the opposite direction to the classical dideoxy chain termination method. Oligonucleotide complementary to the 5'-end of the multipurpose cloning sequence, with the proper restriction enzyme, is used to cleave specifically the molecules to expose a unique 5'-end, upstream to the inserted DNA, for the kinase labeling reaction. No further treatments are necessary before Maxam-Gilbert chemical sequencing reactions.
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004262:DNA Restriction Enzymes; D004277:DNA, Single-Stranded / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel
D004277:DNA, Single-Stranded; D004262:DNA Restriction Enzymes