Patent ID: 7820412

Claim:
A method for using consensus shuffling to produce an error-reduced double stranded DNA target sequence, the method comprising the steps of: (a) providing at least one double stranded DNA molecule having a predefined target sequence, wherein the predefined target sequence has at least one error in said target sequence, and wherein the error comprises a sequence mismatch between the duplex strands; (b) digesting the double stranded DNA molecule of step (a) with at least two endonucleases into smaller overlapping duplex DNA molecules, such that at least one of the smaller overlapping duplex DNA molecules contains a sequence mismatch between its duplex strands; (c) exposing the digested smaller duplex DNA molecules to a DNA binding agent which binds selectively to duplex DNA molecules having a sequence mismatch between their duplex strands for the purpose of removing the smaller duplex DNA molecules having the sequence mismatch; (d) separating the digested and bound smaller duplex DNA molecules having a sequence mismatch away from the digested smaller duplex DNA molecules of correct sequence, wherein the remaining smaller duplex DNA molecules are enriched in the correct target sequence; and (e) assembling the double stranded DNA molecule having an error-reduced target sequence by hybridizing the overlapping smaller duplex DNA molecules of step (d) which are enriched for the correct target sequence to each other, and using the resulting hybridized DNA molecules as templates in PCR to amplify the enriched DNA molecules into the double stranded DNA molecule having an error-reduced target sequence.