A new strategy to create ordered deletions for rapid nucleotide sequencing.
A method is described for generating ordered deletions using previously published techniques but a new strategy. This method is simpler than the published ones and has many advantages. Target DNA is cloned in both orientations into one of the unique restriction enzyme sites adjacent to the complementary region of the commercially available primers in bacteriophage M13. Ordered unidirectional deletions are created using BAL 31 nuclease and religating into M13 vector DNA without the need of purifying BAL 31-digested DNA from a gel.
D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D003090:Coliphages / Q000235:genetics; D004706:Endodeoxyribonucleases; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008967:Molecular Biology / Q000379:methods*
D004706:Endodeoxyribonucleases; C038407:exonuclease Bal 31