KBC/MBIO Practicals in Molecular Biology

Guarantor: Petr Galuszka
Tutorial lecturers: Ondřej Plíhal, David Zalabák
Exercise: 100 hours/semester
Credits: 7
Summer semester
Form of course completion: course credit

• Inoculation and cultivation of E. coli cells bearing selected plasmids. Isolation of plasmid DNA from bacterial cells. Restriction cleavage of plasmid DNA. Agarose electrophoresis.  DNA quantitation.
• Detection of recombinant DNA in food by PCR. Isolation of DNA from flour. Setup and evaluation of the PCR reaction.
• Transformation of yeast S. cerevisiea using a shuttle vector and integration vector bearing the gene Atckx2. Selection of transformants and comparison of transformation yield. Evaluation by measuring CKX activity.
• Work with bacteriophage cDNA library. Cultivation of the phage, DNA isolation. Determination of the titre of the library. Detection of a low abundant and a house-keeping gene.
• Isolation of RNA from plant material, reverse transcription and RT PCR.