Heteroduplexes with mismatches at four sites were constructed from separated strands of lambda DNA and used to transfect Escherichia coli under recombinationless conditions. The output phages from 967 single cells in one experiment and 1016 in another were analyzed to determine the pattern of mismatch repair. A wide range of repair frequencies was found among the mismatches studied. Repair involving two or more close sites in the same heteroduplex occurs much more often on thesamestrandthanonopposite strands. Analysis of the pattern of repair suggeststhat repair tracts initiate at mismatches, propagate preferentially in the 5'leads to 3' direction, and extend an average distance of ca 3000 nucleotides.