Recombinant DNA Technology PPTs

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: M13 phage vectors phagmid PPT  

Lambda phage vectors cosmid PPT

Yeast cloning vectors, Cloning vectors for plants and animals. Artificial chromosomes BAC, PAC & YAC.

Isolation of nucleic acid from bacteria, animal, plants, plasmid DNA and Phage DNA.Formation of DNA fragments using linkers adaptors and homopolymer tails

Cloning strategies  (cell based cloning and PCR based gene cloning

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.