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Identification of a chromosomal mutation by PCR. Chromosomal DNA was isolated from transconjugants or wild-type cells and was used as a template for PCR with primers corresponding to the 5′ and 3′ ends of impA. Products were analyzed by agarose gel electrophoresis. Lane A, 1-kb DNA ladder; lane B, wild-type chromosomal DNA; lane C, cloned impA disrupted with a spectinomycin cassette; lane D, impA mutant chromosomal DNA.
