Quantitation and Purification of Polymerase Chain Reaction Products by High-Performance Liquid Chromatography

Abstract

The polymerase chain reaction (PCR) has rapidly become a standard laboratory technique. With the continuous development of PCR technology there is now a growing need for PCR product quantitation in areas such as therapeutic monitoring and quality control, disease diagnosis, and regulation of gene expression. One of the most common methods currently employed for post-PCR analysis is agarose or poly-acrylamide gel electrophoresis. However, the method is time-consuming and only semiquantitative. In contrast, high-performance liquid chromatography (HPLC) is well accepted as a quantitative technique in many diverse applications areas such as pharmaceutical, biotechnology, food, and environmental, since the technique can provide reliable, precise, and sensitive sample detection, and wide dynamic range.