The book deals with some of the important techniques of recombinant DNA technology including ; isolation of plasmids and transformation of bacteria, isolation and characterization of nucleic acids, construction and screening of gene libraries, DNA sequencing, RFLP and RAPD gene mapping, genetic transformation and molecular analysis of the transgenic plants.
The book includes protocols of some the important techniques currently in use in the field of plant biotechnology. The book also includes methods used in RFPL and RAPD analysis and a practical example of actual research problem in the field of plant genetic engineering. A glossary of important terms is also included which will be helpful to understand the subject matter. The preparation of common buffers and reagents used in the different protocols is included in the appendix.
1. Background - Plant Biotechnology, 2. Isolation, Restriction and Agarose Gel Electrophoresis of E.coli Plasmid DNA, 3. Transformation of Bacterial Cells , 4. Preparation of Bacterophage? DNA ,5. Isolation and Purification of Genomic DNA from Plants, 6. Isolation of Total RNA and in vitro Translation of Poly (A)+RNA, 7. Isolation of and Analysis of Polyribosomes and Ribosomal Proteins,8. Preparation of a Probe: Radioactive Labeling of Nucleic Acids, 9. Southern and Northern Blotting and Hybridization, 10. Construction of Genomic Library, 11. Screening of Genomic Library, 12. Construction and Screening of cDNA Library, 13. PCR and RT-PCR Methods, 14. M13 Cloning for Di-deoxy DNA Squencing, 15. DNA Sequencing Methods, 16. Isolation and in vitro Culture of Protoplasts, 17. Methods of Genetic Transformation of Plants, 18. Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorphic DNA (RAPD) Analysis, 19. Development and Characterization of Transgenic Crop Plants