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colonies, while the addition of the methylated BamHl fragment resulted in 22 transformants. The presence of an equal amount of nonmethylated BamHl fragment stimulated the number of transformants to 335. This 15-fold stimulation of transformatio n frequency is attributed to the absence of the methyl group at the A residues of the BamHl ends, since neither of the BamHl fragments in the absence of vector contributed to the number of transformants. Mini DNA preps were performed on six of the damr and six of the dam+ transformants. Restriction digests demonstrated that all of the damr transformants contained plasmids that acquired the BamHl fragments, while 2 of the 6 damr clones had recombined to form aberrant plasmids. The inhibition of ligation of 2 base overhangs by methyl groups has been reported previously and was found to be the reason that Pvul ends ligate to each other at low frequencies (New England Biolabs 1988-1989 catalog, p. 27). We speculate that ligation of the partially filled cohesive ends created by any restriction enzyme that cuts at dam or dcm sites will be inhibited by the methyl groups. Because many enzymes that can be used for partial fill-in ligations, such as Bell and Bgll, cut at dam methylation sites, we recommend that DNA for these purposes be isolated from dam~ strains.

The absence of adenine methylation increases the ligation efficiency of partially filled staggered DNA ends