The course covers PCR-based DNA/RNA analysis from sampling to evaluation of results, with a specific focus on real-time quantitative PCR (qPCR). The aim of the course is to improve the understanding of the biochemical and physical principles that govern PCR, thus giving the participants the tools for designing, optimizing, validating, trouble-shooting and using PCR/qPCR assays.
Included topics are (i) absolute and relative quantification of DNA/RNA, (ii) limit of detection, (iii) optimization of PCR, (iv) primer/probe design, (v) contamination and quality control, (vi) validation of PCR methods (vii) sample preparation (DNA/RNA), (viii) understanding and relieving PCR inhibition (pre-PCR processing), and (ix) forensic DNA analysis.
Laboratory exercises are integrated with the lectures and serve to illustrate essential theoretical aspects of PCR design and optimization. The obtained results will be thoroughly discussed within the course. Additionally, various PCR-based methods and techniques will be presented and discussed, including (i) gene expression (ii) digital PCR, (iii) high resolution melting (HRM), (iv) and massively parallel sequencing (MPS). Finally, all students will give an oral literature presentation on a chosen topic related to PCR methodology. Course literature, lab manuals and lecture slides will be provided on the first day of the course.
The course leader performs research in the areas of diagnostic PCR, forensic DNA analysis, and pre-PCR processing. The course was started by Professor Peter Rådström in 1997 and real-time quantitative PCR has been applied in the course since 1999.
Adjunct Associate Professor, Division of Applied Microbiology
Faculty of Technology (LTH)
Specialist, National Forensic Centre
Swedish Police Authority