Homogeneous Fluorescent Chemistries for Real-Time PCR
M. A. Lee, D. J. Squirrell, D. L. Leslie and T. Brown
from: Real-Time PCR: Current Technology and Applications (Edited by: Julie Logan, Kirstin Edwards and Nick Saunders). Caister Academic Press, U.K. (2009)
Abstract
The development of fluorescent methods for a closed tube polymerase chain reaction has greatly simplified the process of nucleic acid quantification. Current approaches use fluorescent probes that interact with the amplification products during the PCR allowing kinetic measurement of product accumulation. These probe methods include generic approaches to DNA quantification such as fluorescent DNA binding dyes. There are also a number of strand-specific probes that use the phenomenon of Fluorescent Energy Transfer. In this chapter we describe these methods in detail, outline the principles of each process, and describe published examples. This text has been written to provide an impartial overview of the utility of different assays and to show how they may be used on various commercially available thermal cyclers read more ...
