Techniques of Molecular Biology and Biotechnology

1. Techniques of Molecular Biology and Biotechnology http://powerpointpresentationon.blogspot.com

2. Contents • Introduction • Materials and Method • Observation and results • Discussion

3. AIM • SDS PAGE Analysis • Transformation • Plasmid DNA isolation

4. Introduction • Among biochemical techniques SDS-PAGE are widely used due to its simplicity and effectiveness for describing the genetic structure of crop germplasm. • SDS-PAGE can be used as a promising tool for distinguishing cultivars of particular crop species. • Transformation is genetic alteration of cell and expression of exogenous DNA from its surrounding and taken up through cell membrane. • Isolation of plasmid DNA from E.Coli is a common routine in recombinant DNA technology.

5. Material and Method • SDS PAGE:- • Materials required:- Seeds, Stock solution, Staining solution, Destaining solution, electrode assembly etc. • Procedure:-Gel prepared without any disturbance. • Transformation:- • Requirements:-Centrifuge, water bath, E.coli culture etc. • Procedure:-Take competent cells ,thaw it on ice,add plasmid DNA, give heat shock , plate it on selective media. Cells counting plasmid DNA form colonies.

6. Plasmid DNA isolation:- • Materials Required:-Solution A,B,C, ice, centrifuge etc. • Procedure:-Grow culture , collect supernatant, add solutions, incubate, centrifuge.

7. Observation and Results • SDS PAGE:- Bands were formed at different molecular weight for different varieties of wheat.

8. Bacterial Transformation:-Blue colonies indicating transformed colonies and white indicating colonies without plasmid.

9. Plasmid DNA isolation

10. Conclusion • Process of screening bacterial transformants for recombinant plasmids is more rapid and simple by use of vectors. • SDS PAGE detect presence of protein. • After plasmid isolation the DNA isolated was pure enough to be used for cloning. It separate plasmid DNA from cellular debris and chromosomal DNA in pellet.

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