Caister Academic Press

Internal and External Controls for Reagent Validation

M. A. Lee, D. L. Leslie and D. J. Squirrell
from: Real-Time PCR: Current Technology and Applications (Edited by: Julie Logan, Kirstin Edwards and Nick Saunders). Caister Academic Press, U.K. (2009)

Abstract

False negatives in PCR can occur from inhibition of one or more of the reaction components by a range of factors. Therefore applications requiring a high level of confidence in the result need to be designed to control for the occurrence of false negatives. While an external, or batch, control is often used, the ideal control is one that is included in the reaction cocktail in a multiplex format. Here we discuss the application and development of molecular mimics for use as controls in real-time PCR, and explain a number of concepts and experimental considerations that will aid in the optimisation of controlled multiplexed assays read more ...
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