DNA sequencing with direct blotting electrophoresis.
A method for transferring the DNA molecules of sequencing reaction mixtures onto an immobilizing matrix during electrophoresis has been developed. A blotting membrane moves with constant speed across the end of a very short, denaturing gel and collects the molecules according to size. A constant distance between bands for molecules differing in length by one nucleotide is obtained over a large range (approximately 600 nucleotides with a 5% gel), simplifying the determination of DNA sequences considerably. Reliable sequences of 500 nucleotides can be read and sequence features up to greater than 1000 nucleotides are revealed in a single experiment. The sequencing of a potential Z-DNA-forming fragment from Escherichia coli DNA is given as an example and possible further developments are discussed.
D001483:Base Sequence; D004247:DNA; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli; D010957:Plasmids
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004247:DNA