Abstract
Publisher Summary In vitro polymerase chain reaction (PCR)-based methods for recombining homologous deoxyribonucleic acid (DNA) sequences are capable of creating highly mosaic chimeric sequences. Several different methods have been reported for in vitro recombination or DNA shuffling: for example, the original Stemmer method of DNase I fragmentation and reassembly, the staggered extension process (STEP), and random priming recombination. Slight variations in the shuffling protocols can affect the outcome of the experiment. Different gene sequences recombine most efficiently under different conditions. This chapter provides protocols that are designed to give a high likelihood of success. The protocols are known to work for recombining sequences of ∼>85% identity.
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