Coliphage M13 cloning system and ddXTP chain-termination method for DNA sequencing.
Two DNA fragments of cytochrome b gene in yeast mitochondrial DNA were sequenced by Messing's M13 cloning system and Sanger's ddXTP chain-termination method. M13mp8 and M13mp9 serve as vectors for insert fragment. The recipient strain is E. coli JM103 for transfection. When the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. The two fragments, which have 575 bp and 709 bp, were sequenced. To read more bases, the ratio of ddXTP/dXTP must be reduced. On one X-ray film of 80 x 17 cm 394 bp were read.
D001483:Base Sequence; D003090:Coliphages / Q000235:genetics*; D004251:DNA Transposable Elements; D004272:DNA, Mitochondrial / Q000235:genetics; D004274:DNA, Recombinant; D005822:Genetic Vectors; D015003:Yeasts / Q000235:genetics
D004251:DNA Transposable Elements; D004272:DNA, Mitochondrial; D004274:DNA, Recombinant