Kinetics of competitive reverse transcriptase-PCR

Publisher Summary Competitive reverse transcriptase (RT)- polymerase chain reaction (PCR) is a versatile tool for quantification of gene expression. It is a technique whose past has been shaded by doubts and questions concerning its reliability. However, these suspicions have been subsided with time. And with the coming of new analytical tools, it has been possible to explore biological function as manifested in variations in the level of gene expression. This chapter discusses the kinetics of competitive RT-PCR. The reason why gene expressions need to be measured is the fact that when competitive RT-PCR methods are carefully designed and applied, both precision and accuracy are high, even when microscopic tissue samples are analyzed. The chapter also describes the applications of quantitative gene expression. This approach is useful in two general arenas – clinical laboratory and research laboratory. The several advantages of using these approaches in these two areas are discussed in this chapter. It also unravels the critical elements of performance in competitive RT-PCR. Finally, the performance characteristics are discussed that include versatility, construction, sensitivity, and validation.