Creation of Chimeric Junctions, Deletions, and Insertions by PCR

Recombinant polymerase chain reaction (PCR) (1 ) is the method of choice if one wants to modify a cloned DNA. It is a versatile technique that allows operations as different as creation of deletions, addition of small insertions, site-directed mutagenesis, and construction of chimeric molecules at any chosen location in the molecule of interest (see Note 1 ). This chapter describes in detail a simplification of the original recombinant PCR method. This fast and efficient method has been successful in fusing two different sequences with precision (2 –4 ). It can also be used to create deletions or insert small fragments of DNA.