Recombinant DNA Technology PPTs

Introduction to Recombinant DNA technology and Gene Cloning

Restriction endonucleases and DNA modifying enzymes (ligase, polymerases, alkaline phosphatase, polynucleotide kinase, terminal transferase, Reverse transcriptase)

Cloning vectors:

Plasmids,

Phages,

Cosmids,

Yeast cloning vectors,

Animal and plant viruses as vectors. Artificial chromosomes

BAC, PAC & YAC.

Isolation of nucleic acid from bacteria (plasmid and genomic DNA), Plant, animal & bacteria.

Basic steps of gene cloning

Cloning strategies  (cell based cloning and PCR based gene clonoing

Genomic library

Synthesis of cDNA. Construction of cDNA library

Selection of r DNA clones and their expression products, Chromosome walking.

Expression of cloned genes in heterologous host.

DNA sequencing: Chemical and enzymatic methods.                                                             0.5

PCR: Types and applications. Real Time PCR. Site directed mutagenesis. Ribonuclease protection assay, Gel retardation assay,

DNA foot printing, DNA finger printing, DNA profiling.

Genomic analysis: S-1 mapping, RFLP, RAPD, AFLP.                                                           0.5

Probe labeling and hybridization. Blotting techniques: Southern, Northern and Western blotting (Methodologies and applications)

Transgenic Technology: Types, approaches & applications (Plant & Animals),                     0.5

Gene therapy: Principles, strategies and ethics of gene therapy. Genome editing technologies: Principles and applications.