SUBJECT

Molecular genetics

practical

master

Faculty of Science

Biology MSc

4

Semester 2

Spring semester

In 4 separate blocks.

Block I (Days 1-3): Gene silencing (RNA interference) in genetic analysis. In silico (bioinformatics) design of primers, total RNA isolation, mRNA isolation, RT-PCR (reverse transcriptase PCR), cloning of a cDNA fragment into a „feeding” vektor (digestion, fragment purification, ligation), bacterial transformation. Phenotypic (phenocopy) analysis. Analysis of genetic pathway.

Block II (Days 4-6): Molecular techniques in genetic analysis I: PCR, Southern Blotting (detection of insertional, deletional and point mutations), Northern analysis (hybridization of ceh-20/bec-1 mRNA probes to RNA extracts from different developmental stages – developmental Northern).

Block III (Days 7-8): Western blotting (Detection of BEC-1 protein in different cell farctions – nucleus, organellum, cytoplasm). Genotyping with microsatellite markers.

Block IV (Days 9-10): Determining individual DNA patterns, sex determination.

• Anthony J.F. Griffiths, Susan R. Wessler, Sean B. Carroll, John Doebley "An introduction to genetic analysis”, 10th edition, W. H. Freeman, 2010

• SF Gilbert "Developmental Biology”, 7th edition, Palgrave Macmillan, 2003

• EH Davidson "Genomic regulatory systems”, Academic Press 2002

• B Lewin "Genes”, Jones & Bartlett Learning 2005