The next four chapters cover DNA extraction, DNA quantification, factors affecting PCR reliability and validity, and quantitative realtime PCR. The third chapter introduces the general principles of method validation and the concepts of accuracy, precision, limit of detection, limit of quantification, measurement uncertainty, etc., and then gives a worked example of the validation of a realtime PCR procedure. The second chapter discusses quality in the molecular laboratory and provides pointers to the relevant ISO and other standards, and stresses the need for sample and metrological traceability. The first chapter sets the context for the rest of the book, introducing ‘Valid Analytical Measurement’ principles, which will be familiar to those working in ISO17025 and similar environments. This book takes the welcome approach of focussing on quality control, quality assurance, optimization, validation and calibration. Given that nucleic acid analyses are now used routinely in forensic and food safety fields, robust methods are highly desirable, if not essential. There are many books available that introduce the uninitiated to the delights of nucleic acid analysis, mostly in the form of recipe collections, and such books succeed or fail by the attention to detail given to optimization, standardization and trouble-shooting protocols. The analysis of the sequence, diversity and abundance of nucleic acids underpins modern biology.
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