Course

Unit 1

Introduction to rDNA technology

The Basic Principles of Gene Cloning and DNA Analysis   Introduction, History, the advent and importance of gene cloning and the polymerase chain reaction, Vectors for Gene Cloning, Purification of DNA from Living Cells, Manipulation of Purified DNA, Introduction of DNA into Living Cells

Unit 2

Vectors for Cloning

Cloning Vectors for E. coli, λ and other high-capacity vectors, Cloning Vectors for Eukaryotes, Genomics & cDNA Libraries

Unit 3

Applications and Techniques of Gene Cloning   

Polymerase Chain Reaction & qPCR, Electrophoresis & Blotting Techniques, Site- Directed Mutagenesis, DNA Sequencing, Reporter Gene Assays, DNA-Protein Interaction Assays, Protein-Protein Interaction Assays, DNA Fingerprinting.

LEARNING OBJECTIVES:

The course attempts to introduce the basic concepts of recombinant DNA technology namely Boyer and Cohen’s workflow of gene manipulation, restriction and ligation, plasmid and phage-based vectors, transformation techniques, site-directed mutagenesis and applications. 

COURSE OUTCOMES:

After completing the course, students shall be able to

CO1. Describe basic concept of cloning, various steps involved, tools used like restriction enzymes, PCR, primer designing, vectors (Knowledge).  

CO2. Interpret & discuss the principles of different molecular tools like  qPCR, DNA sequencing, Blotting electrophoresis, reporter assay, DNA fingerprinting, DNA-protein and protein-protein interactions (Understand).  

CO3. Illustrate the knowledge to apply for recombinant protein production,  transgenic plant production for use in agriculture, medicine and research (Apply).

REFERENCES:

1. A. Brown, Gene Cloning and DNA Analysis: An Introduction, 6th Edition, Wiley-Blackwell.
2. Sandy B. Primrose, Richard Twyman, Principles of Gene Manipulation & Genomics – 7th Edition –– Blackwell