Practical Biotechnology and Plant Tissue Culture

Contents
1. Principle, working and uses of instruments: Autoclave, Hot air oven, Incubator, Laminar air flow, pH meter. Spectrophotometer, Colarimeter, Centrifuge, Ultracentrifuge, Paper chromatography, Thin Layer chromatography, Paper electrophoresis, Gel electrophoresis
2. Preparation of culture media
3. Preparation of standard solution
4. Isolation of bacteria by streak plant method
5. Isolation of bacteria by pour plate method
6. To determine the growth characteristics of E.coli using plating technique
7. Growth characteristics of E.coli using turbidimetric method
8. Isolation of plasmid from E.coli by alkali lysis method and to analyze plasmid DNA by agarose gel electrophoresis
9. Isolation of bacteria genomic DNA from E.coli or Serratia marsences
10. To determine the DNA molecular size
11. Purification of DNA Molecular size
12. To perform Bacteriophage titration
13. To perform restriction digestion of DNA with enzyme viz. Eco RI and Hind III and to analyze the restriction pattern by agarose gel electrophoresis
14. To perform transformation of the given bacterial population
15. DNA amplification by Polymerase chain reaction
16. Preparation of Plant Tissue culture media
Preparation of Explant and its sterilization
17. To isolate protoplast from the plant tissue
18. Viability testing of protoplast
19. Demonstration of protoplast fusion using polyethylene glycol
20. Demonstration of Callus culture technique
21. Initiation of callus from radical tip of Phaseolus vulgaris
22. Demonstration of cell suspension culture
23. Demonstration of Androgenesis is Dature
24. Demonstration of Somatic embrogenesis
25. Demonstration of Organogenesis
26. Estimation of Growth Index
27. Preparation of Artificial seeds
28. Transfer of Plantlets to green house condition/field
29. Isolation of Genomic DNA from plant cells
30. Isolation of Agrobacterium sps. From soil & study of growth characteristics
31. Isolation of Agro bacterium sps. From crown gall
32. Demonstration of b-glucuronidase (gus) activity by histrochemical assay
33. Demonstration of DNA polymorphism through Restriction Fragment Length Polymorphism (RFLP) of different crop varieties
34. Demonstration of DNA polymorphism through Random Amplified Polymorphism DNA (RAPD)