TRAP-Mutant™ DNA Shuffling Kit

TRAP-Mutant™ DNA Shuffling Kit is an excellent tool for molecular evolution of genes. It is based on DNAse-I random fragmentation of one target or a collection of homologous genes. In vitro homologous recombination of fragment pool is generated by a primerless PCR. These fragments are reassembled into a full-length gene by repeated cycles of annealing in the presence of DNA polymerase.

Reassemble of one shuffled gene generates point mutations at a relatively high rate (0.7%) while the reassemble of several shuffled genes generates a library of chimeric genes with point mutations. A single cycle of shuffling yielded 8 fold improvements from four separately evolved genes, versus a 270- to 540-fold improvement from the four genes shuffled together, a 50-fold increase per cycle of shuffling.