Horizon has launched a book that discusses the strategies for preparing effective controls and standards for PCR, when they should be employed and how to interpret the information they provide.
The book, by Suzanne Kennedy and Nick Oswald (MO Bio Laboratories, Carlsbad, California, US; and Bitesize Bio, Edinburgh, UK, respectively) highlights the significance of optimisation for efficiency, precision and sensitivity of PCR methodology and provides guidance on how to troubleshoot inefficient reactions.
Experts in PCR describe design and optimisation techniques, discuss the use of appropriate controls, explain the significance of standard curves, and explore the principles and strategies required for effective troubleshooting.
Authors highlight the importance of sample preparation and quality, primer design, controlling inhibitors, avoiding amplicon and environmental contamination, optimising reagent quality and concentration, and modifying the thermal cycling protocol for optimal sensitivity and specificity.
Specific chapters discuss the history of PCR, the choice of instrumentation, the applications of PCR in metagenomics, high-resolution melting analysis, the MIQE guidelines and PCR at the microlitre scale.
The strategies, tips and advice included enable the scientist to optimise and effectively troubleshoot a range of techniques, including PCR, reverse transcriptase PCR, real-time PCR and quantitative PCR.
