A facile DNA shuffling protocol was introduced. Two genes of about 1 700 bp were obtained by PCR from two different individual templates, separately, which shared over 93% homology. After mixed with equimolar of each, these two genes were further cut randomly by DNaseⅠ into small fragments of 10～50 bp under the existence of Mg²⁺. These small fragments were successfully re-assembled to form full genes with original size by one round of PCR without any external primers and two other rounds of normal PCR amplification. This shuffling protocol may help to construct chimera genes from a family of genes with high sequence homology.