Direct Sequencing of PCR Products with DNA-Binding Proteins

Abstract

The polymerase chain reaction (PCR) has become widely established as a powerful core molecular biology technique because of its ability of produce large amounts of specific target DNA from limited template sources (1). Numerous applications based on the PCR have also been developed, including site-directed mutagenesis, in vitro expression, and nucleotide sequencing (2,3). The combination of PCR amplification and direct sequencing of products is a highly desirable procedure that, in many cases, obviates the need for complex and time consuming cloning protocols (4). However, there appears to be no one universally accepted method for direct PCR sequencing (5).