Technical Notes for the Detection of Nucleic Acids

Abstract

In following any polymerase chain reaction (PCR)-based method, it is usual to identify the products of the reaction by some form of detection system. The majority of these still rely on size- and charge-based separation systems, although for some quantitative PCR applications, either direct measurement of fuorescence or indirect Enzyme-linked immunosorbent assay-based systems can be used. In this chapter, we summarize some of the most common methods for detection of nucleic acids as a handy reference for those seeking to validate their PCR reactions. Although there are many variants of these techniques, we have confined our reporting to methods of which we have direct experience and which offer broad applicability. Fluorescence detection of quantitative real-time PCR requires specialist equipment, and we have therefore omitted this from our discussions at present.