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)
CRISPR-associated nucleases (Cas9 Cas12 Cas13) PPTx
Taq Pfu Phusion DNA polymerase
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.
Introduction of DNA into Host Cells (Bacteria, Plant and Animal cells)
Selection and Scorable markers/Reporter genes
Cloning strategies (cell based cloning and PCR based gene cloning)
Synthesis of cDNA. Construction of cDNA library
Methods of identification of clone from gene library (library screening strategies)
Expression of Cloned Gene in Heterologous System: Prokaryotes ;
Basic architecture of an expression vector; Critical components of an expression vector; Expression of fusion protein
Expression of cloned genes in heterologous host (Fungi, Insects
Screening and Characterization of Recombinant Proteins PPT
BLOTTING TECHNIQUES: Southern, Northern, Western, Dot, Slot, and Far-Western blotting
Probe labeling: Radioactive and Non-radioactive PPT
Principle of nucleic acid hybridization
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.
Transgenic Technology: Types, approaches & applications (Plant & Animals), 0.5
Gene therapy: Principles, strategies and ethics of gene therapy. Genome editing technologies: Principles and applications.