Creation of Chimeric Junctions, Deletions, and Insertions by PCR

Abstract

Recombinant polymerase chain reaction (PCR) () 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 (, , ). It can also be used to create deletions or insert small fragments of DNA.