Quality Control of Nucleotides and Primers for PCR

The polymerase chain reaction (PCR) has become an essential tool in molecular biology for the rapid amplification and analysis of DNA (1 ). The technique requires the use of oligonucleotide primers that bind to the DNA at a specific region of interest. The region is subsequently copied by enzymatic polymerization of deoxynucleotides. Among its many applications, PCR has found use in fields where the quality of results is of primary importance, such as in disease diagnosis (2 ) or forensic DNA analysis (3 ). In these cases, PCR amplification from very small quantities of DNA is often required (4 ,5 ), requiring high-reaction efficiency. Additionally, the consequences of ambiguous or false results in assays are far-reaching, and it is therefore essential that the PCR is highly specific. In order to obtain such high-performance standards the components of the PCR must be carefully controlled, particularly with respect to deoxynucleoside triphosphates (dNTPs) and oligonucleotide primers.