QPCR: Target Preparation
Tania Nolan, Reinhold Mueller and Stephen Bustin
from: Real-Time PCR in Microbiology: From Diagnosis to Characterization (Edited by: Ian M. Mackay). Caister Academic Press, U.K. (2007)
Application of modern molecular biology techniques, for the detection of pathogens has resulted in a radical change in the methodology adopted by many microbiology laboratories for the diagnosis of pathogens. Use of real-time quantitative PCR technology has resulted in much more sensitive and specific detection. It is becoming apparent that the most vulnerable stages of the qPCR and RT-qPCR protocols are those concerned with target preparation. Each specimen, organism and nucleic acid combination requires a different set of considerations for collection, transport and nucleic acid purification and the most appropriate combination should be ascertained for each project and laboratory. In order to ensure that the data collected are reliable the relevant controls should also be included. In this way a set of standard operating procedures can be defined that ensure reliable, reproducible and robust data read more ...