Molecular Biology Compact Course

Target Group

Technologists or scientists without any previous knowledge or possessing only basic knowledge

Content/Learning Objective

The course will give an introduction into standard molecular biology techniques. The aim is to clone an interesting region of the bacteriophage Lambda into the plasmid vector pUC18. The properties of DNA and protein, their isolation, production and manipulation are taught theoretically as well as practically. Starting with genomic DNA, we will amplify an interesting gene fragment using polymerase chain reaction (PCR), digest it with restriction enzymes and insert it into the vector (ligation). Additional methods include: agarose gel electrophoresis, transformation, blue white selection, selection and validation of the clones. Then we will focus on protein biochemistry and examine protein expression of the marker gene GFP (green fluorescent protein) that will be isolated from E.coli culture by affinity chromatography. Homogeneity and yield is then checked by SDS polyacrylamide gel electrophoresis and a Bradford protein assay. Finally we will deal with immunomethods and determine the antibody titer of an unknown sample by means of the ELISA technique.

Main Topics

Properties of DNA and protein, their isolation, production and manipulation