Cloning PCR Products with T-Vectors

Abstract

Since it was described in 1988 (1), the polymerase chain reaction (PCR) has been a valuable tool for molecular biologists. PCR allows researchers to produce a large quantity of a desired DNA fragment while requiring only a small amount of template. Prior to PCR, isolation of DNA fragments was typically performed by cleavage of the DNA with restriction endonuclease enzymes. The relative abundance or scarcity of appropriate restriction sites within the region of interest greatly affected the ability of researchers to obtain specific DNA fragments. PCR gives researchers the ability to establish the terminal sequences as well as the size of the amplified fragment and, in this way, provides freedom from the problems associated with location and abundance of restriction enzyme recognition sites.