Sequencing long DNA fragments cloned in bacteriophage M13 by using internal primers. The sequence analysis of a yeast DNA fragment containing a replication origin.
In the ;shotgun' procedure for sequencing DNA, DNA fragments are cloned into a phage M13 vector and sequenced by using a flanking primer. In a variation of this procedure a longer DNA sequence is cloned into M13, the two single-stranded recombinants identified and sequenced by using a set of internal primers prepared by exonuclease III digestion of restriction fragments.
D001483:Base Sequence; D003001:Cloning, Molecular; D003090:Coliphages / Q000378:metabolism*; D004261:DNA Replication; D004271:DNA, Fungal / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000032:analysis; D003851:Deoxyribonucleases / Q000378:metabolism; D005090:Exodeoxyribonucleases; D005092:Exonucleases / Q000378:metabolism; D008722:Methods; D012441:Saccharomyces cerevisiae / Q000032:analysis*
D004271:DNA, Fungal; D004274:DNA, Recombinant; D004277:DNA, Single-Stranded; D003851:Deoxyribonucleases; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C020133:exodeoxyribonuclease III