Department Events
No Upcoming Events
Laboratory IV (Molecular Biology and Genetic Engineering)
The objectives of this course are to provide students with experimental knowledge of molecular biology and genetic engineering
CO107: Determine quality and quantity of isolated plasmid and perform restriction digestion, ligation and valediction of PCR, colony PCR.
CO108: Elaborate the concept of recombinant protein expression, it’s soluble and insoluble forms, purification and verification by SDS PAGE
CO109: Design experiments for gene transfer in prokaryotic cells, Phage titre value and gene expression and it’s regulation
CO110: Identify and illustrate the various elements of molecular biology and genetic engineering
CO111: Discuss and defend the concepts of molecular biology and genetic engineering and illustrate the exercises performed with appropriate methods and outcomes and maintain proper documentation of the same.
CO112: Contribute effectively in course-specific interaction
Molecular Biology and Genetic Engineering
1. Concept of lac-operon: a) Lactose induction of B-galactosidase. b) Glucose Repression. c) Diauxic growth curve of E.coli
2. UV mutagenesis to isolate amino acid auxotroph
3. Phage titre with epsilon phage/M13
4. Genetic Transfer-Conjugation, gene mapping
5. Plasmid DNA isolation and DNA quantitation
6. Restriction Enzyme digestion of plasmid DNA
7. Agarose gel electrophoresis
8. Polymerase Chain Reaction and analysis by agarose gel electrophoresis
9. Vector and Insert Ligation
10. Preparation of competent cells
11. Transformation of E.coli with standard plasmids, Calculation of transformation efficiency 12. Confirmation of the insert by Colony PCR and Restriction mapping
13. Expression of recombinant protein, concept of soluble proteins and inclusion body formation in E.coli, SDS-PAGE analysis
14. Purification of His-Tagged protein on Ni-NTA columns a) Random Primer labeling b) Southern hybridization
