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DNA amplification technology - week 42

Course description
The course covers real-time quantitative PCR (qPCR) analysis from sampling to evaluation of results. The aim of the course is to improve the understanding of the biochemical and physical principles behind PCR, thus giving the students the tools for designing, optimising and using PCR/qPCR assays.

Included topics are (i) absolute and relative quantification of DNA/RNA, (ii) conventional PCR vs. qPCR, (iii) reverse transcription qPCR, (iv) optimisation and kinetics of qPCR (v) primer/probe design, (vi) sample preparation (DNA/RNA), (vii) understanding and relieving PCR inhibition, and (viii) quality assessment of qPCR results.

Laboratory exercises are integrated with the lectures and serve to illustrate essential theoretical aspects of PCR design and kinetics. The obtained results will be thoroughly discussed within the course. Additionally, various applications of PCR/qPCR, such as diagnostic PCR, digital PCR, forensic DNA analysis, gene expression, and high resolution melting, will be presented and discussed. Finally, all students will give an oral presentation on a chosen topic.

The course leaders perform research in the areas of diagnostic PCR, food microbiology, forensic DNA analysis, gene expression, kinetics and modelling of qPCR, and pre-PCR processing. Real-time PCR has been applied in the course since 1999.
Course manual, lecture slides and literature will be provided. More information can be found on the course home page.

Course leaders
Johannes Hedman
Applied Microbiology
Faculty of Technology (LTH)
Lund University
and
Swedish National Laboratory of Forensic Science (SKL), Linköping

Peter Rådström
Applied Microbiology
Faculty of Technology (LTH)
Lund University

More information
Johannes Hedman
Tel: +46 (0)46 222 83 29
E-mail: johannes.hedman@tmb.lth.se

Course home page
http://www.tmb.lth.se/education/courses/dna-amplification-technology/