Practical
- Media preparation, sterilization and inoculation of given material.
- Growth characteristics of E.coli using plating and turbidimetric methods.
- Isolation of plasmid from E.coli by alkaline lysis method and its quantitation spectrophotometrically.
- Restriction digestion of the plasmid and estimation of the size of various DNA fragments.
- Cloning of a DNA fragment in a plasmid vector, transformation of the given bacterial population and selection recombinants.
- Organogenesis and somatic embryogenesis using appropriate explantsand preparation of artificial seeds.
- Isolation of protoplast from various plant tissues and testing their viability.
- Effect of physical and chemical factors protoplat yield.
- Demonstration of protoplast fusion employing PEG.
- Demonstration of DNA sequencing using Sanger,s dideoxy method.
- Demonstration of microtechniques for plant cultures.
- Co-cultivation of the plant material with Agrobacterium and study GUS activity histochemically.
Essential Readings:
- Smith and Roberta H. Plant Tissue Culture: Techniques & experiments.
- Chawla, HS. Plant Biotechnology: Laboratory Manual for Plant Biotechnology.
- Philips, GC. 1995. Plant Cell, Tissue and Organ Culture: Fundamental Methods. Narosa Publishing House