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Indian Institute of Science Education and Research Bhopal

BIO 310: Genetic Engineering and Biotechnology (3)

Prerequisites: BIO 301: Cell Biology , BIO 303: Biochemistry I

1. Basic principles of gene cloning
2. Polymerase chain reaction – Theory and applications: Nested, Inverse, Asymmetric, Hot start, touchdown etc. Troubleshooting. RT-PCR
3. Site-directed mutagenesis and random mutagenesis.
4. Vectors for gene cloning – features, applications.
5. Methods of molecular cloning: strategy design and screening.
6. Usage of restriction enzymes; ligation independent cloning.
7. Construction of DNA libraries and library screening; Reporter gene technology.
8. Gene knockout in bacteria – methods and applications.
9. Bacterial and yeast two hybrid systems – theory and applications.
10. Preparation of biomolecules – isolation and purification.
11. DNA sequencing – capillary based and next gen sequencing.
12. 2-dimensional gel electrophoresis and MALDI based protein identification.

Suggested Books:

• Sambrook and Russell: Molecular Cloning.
• T. A. Brown: Gene cloning and DNA analysis
• Glick, Pasternak and Patten: Molecular Biotechnology

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