We describe here an in vitro method of increasing complementarity in a
heteroduplex polynucleotide sequence. The method uses annealing of
opposite strands to form a polynucleotide duplex with mismatches. The
heteroduplex polynucleotide is combined with an effective amount of
enzymes having strand cleavage activity, 3' to 5' exonuclease activity,
and polymerase activity, and allowing sufficient time for the percentage
of complementarity to be increased within the heteroduplex. Not all
heteroduplex polynucleotides will necessarily have all mismatches
resolved to complementarity. The resulting polynucleotide is optionally
ligated. Several variant polynucleotides result. At sites where either of
the opposite strands has templated recoding in the other strand, the
resulting percent complementarity of the heteroduplex polynucleotide
sequence is increased. The parent polynucleotides need not be cleaved
into fragments prior to annealing heterologous strands. Therefore, no
reassembly is required.