BY4311 Advanced Genetic Engineering Laboratory Syllabus:
BY4311 Advanced Genetic Engineering Laboratory Syllabus – Anna University PG Syllabus Regulation 2021
COURSE OBJECTIVES:
1 Provide hands-on experience in performing basic recombinant DNA techniques.
2 To understand the principle behind each techniques and applications of each methodology in applied biological research.
3 To provide in depth knowledge in creation of recombinant DNA and production of recombinant protein
LIST OF EXPERIMENTS
1. Isolation of vector Eg. Plasmid and Isolation of vector Eg. Genomic DNA
2. Electroporation to Yeast/ Bacteria
3. Isolation of RNA and cDNA synthesis
4. Primer designing and Calculation of Annealing Temperature
5. Demo on Real-time PCR/ Gradient PCR
6. Plasmid isolation and confirming recombinant by PCR and RE digestion.
7. Confirmation of the presence of insert by colony PCR
8. Induction and expression of recombinant protein
9. Western blot with ECL detection
10. Site directed mutagenesis using PCR
11. Southern blot (Non-radioactive)
12. RFLP analysis of the recombinant DNA
13. SDS-PAGE analysis of recombinant protein expression
14. Quantification of expressed recombinant protein by ELISA
LIST OF EQUIPMENTS REQUIRED
Microscopes
PCR
Purification columns
Microplate reader
UV spectrometer
PAGE apparatus,
Western blot apparatus (dry/semi-dry/wet),
Southern blot apparatus,
centrifuge,
Hemocytometer,
CHEMICALS : required stains, chemicals, enzymes & consumables
TOTAL : 90 PERIODS
COURSE OUTCOMES
After completion of the course the students will be able to
CO1 Gain hands-on experience in performing basic recombinant DNA techniques.
CO2 Understand the principle behind each techniques and applications of each methodology in applied biological research.
CO3 Gain depth knowledge in creation of recombinant DNA and production of recombinant protein
REFERENCES
1. Sambrook, J. and Russsel, D.W., “Molecular cloning – A laboratory manual”, Third edition, Cold Spring Harbor Laboratory Press, Cold Spring harbor, New York, USA, 2001