This course will enable the student to grasp plant molecular biology intricacies, harness recombinant DNA techniques and learn tissue culture techniques.
Introduction to PTC laboratory. Sterilization techniques. Plant Tissue Culture media. Concept of totipotency, Embryogenesis and organogenesis and their practical applications. Somaclonal variations and applications.
Callus culture. Embryo culture and its applications, embryo rescue. Micropropagation, axillary bud, shoot- tip and meristem culture to produce virus free plants. Anther culture, Pollen culture. Single cell and suspension cultures.
Protoplast isolation: Principles and applications. Testing of viability of isolated protoplasts. Various steps in the regeneration of protoplasts. Somatic hybridization. Practical applications of somatic hybridization (Hybrids vs. Cybrids)
Gene cloning vectors- Plasmids, Bacteriophages, Cosmids, phagemids, YAC, BAC; Enzymes used in gene manipulation- Restriction endonucleases, DNA polymerase, Polynucleotide kinase, ligase, Terminal deoxynucleotidyl transferase.
Methods of gene transfer, Making and applications of genomic and c-DNA libraries; Blotting Techniques- Southern, Northern and Western Blotting.