Patent ID: 8728734
Filing Date: 2014-05-20
Classification: C12N

Abstract:
1. A method for producing a diversified nucleotide sequence by PCR, comprising; i) performing an error prone PCR reaction on the whole of nucleotide sequence S, wherein said sequence is delimited at its 5′ and 3′ ends by two segments F1 and F2, using at least two primers, one of said primers is a sense strand and the other an antisense strand of segments F1 and F2, respectively, so as to amplify sequence S and introduce random mutations; ii) purifying the amplification products obtained in stage i); iii) performing a high-fidelity PCR reaction starting from the amplification products purified in stage ii), using as sense and antisense primer pairs at least: wherein the primers corresponding to segment D iv) purifying the amplification products obtained in stage iii); v) performing a high-fidelity PCR reaction starting from the amplification products purified in stage iv), using at least one pair of sense and antisense primers F1 and F2; and vi) purifying the amplification products obtained in stage v), wherein the identity of at least N inner domains D