Department Events
No Upcoming Events
LABORATORY IX
|
Course Outcomes |
Learning and teaching strategies |
Assessment Strategies |
|
Upon completion of the course the learner will: CO103: Analyze basic techniques of genomics and proteomics CO104: Clone a gene in bacterial cell CO105: Express protein of interest using an expression system. CO106: Interpret the purification of proteins CO107: Evaluate gene transfer by conjugation |
Interactive Lectures, Discussion, Tutorials, Demonstrations, Assignments Reading Journals |
Class test, Semester end examinations, Quiz, Solving problems in tutorials, Assignments, Presentation, Individual and group projects |
1. Gene cloning
a) Polymerase Chain Reaction and analysis by agarose gel electrophoresis
b) Plasmid DNA isolation and DNA quantitation
c) Restriction Enzyme digestion of plasmid DNA
d) Vector and Insert Ligation
e) Preparation of competent cells
f) Transformation of E.coli with standard plasmids, Calculation of transformation efficiency
g) Confirmation of the insert by Colony PCR and Restriction mapping
2. Expression of recombinant protein, concept of soluble proteins and inclusion body formation in E.coli, SDS-PAGE analysis
3. Purification of His-Tagged protein on Ni-NTA columns
a) Random Primer labeling
b) Southern hybridization
4. Genetic Transfer-Conjugation, gene mapping
