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Home Books Botany Plant Biotechnology And Molecular Biology: A Laboratory Manual

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Blurb

The book, A Laboratory Manual of Plant Biotechnology and Molecular Biology comprises of workable laboratory protocols for a large number of techniques related to plant biotechnology, genetic engineering and molecular biology. This includes plant cell and tissue culture, callus and suspension culture, anther culture, ovule culture, embryo culture, Cryopreservation, Isolation of Plant protoplasts, Protoplast culture and regeneration, production of somatic hybrids through protoplast fusion, gene transformation using Agrobacterium as vector, direct gene transfer using biolistic gun, Isolation of plant and organells DNA, construction and screening of genomic DNA libraries, Molecular markers like RFLP, RAPD, SCARS and CAPS, DNA sequencing, RNA isolation and northern blotting, Isolation of proteins and western blotting etc. The manual is prepared with the objective to cater the needs of post- graduate students as well as for scientists working in the disciplines of Plant Breeding, Genetics, Botany, Plant physiology, Biochemistry, Plant Biotechnology, Molecular Biology etc. It gives an update on some well established methods and presents reliable protocols.

Preface
Chapter
1. Organisation of Plant Tissue Culture Laboratory
2. Media Preparation, Sterilization and Storage
3. Aseptic Seedlings Production and Sterilization of In vivo Grown Plant Material and Excised Explants
4. Micropropagation
5. Initiation and Maintenance of Callus and Suspension Cultures
6. Ovule Culture in Wheat
7. Anther Culture for Haploid and Dihaploids (DH) Production in Sunflower (Helianthus annuus L) 30
8. Embryo Culture for Crop Improvement
9. Somatic Embryogenesis in Alfalfa (Madicago sativa)
10. Production of Synthetic Seeds
11. In Vitro Selection for Salt Tolerance
12. In Vitro Selection for Disease/Toxin (Verticillium Wilt) Resistance Cell Lines in Alfalfa
13. Cryopreservation for Germplasm Conservation
14. Determination of Endogenous Content of Cytokinins in Plant Material
15. Identification of Viruses and/or Nucleocapsid Protein by ELISA
16. Isolation and Culture of Plant Protoplasts
17. Production of Somatic Hybrids Through Protoplast Fusion
18. Electroporation of Protoplasts
19. Genetic Transformation of Plants by Using Agrobacterium tumefaciens Vector System
20. Agrobacterium tumefaciens Mediated Transformation
21. Assaying Marker Genes in Plants-GUS (β-Glucuronidase) Gene
22. Detection of Neomycin Phosphotransferase II (NPT II) Activity in Transformed Plant Tissues
23. Genetic Transformation of Plants Using Agrobacterium rhizogenes Vector System
24. Tobacco Chloroplast Transformation using Micro Projectile Bombardments (Biolistic Gun)
25. Micro Projectile Mediated Transformation of Rice Using Biolistic Gun
26. Inoculation and Storage of Escherichia coli from Overnight Culture
27. Construction of Genomic DNA Library
28. Isolation of Bacteriophage Lambda DNA
29. Screening of Genomic and cDNA Libraries
30. Isolation of Plasmid DNA (Alkaline Lysis Method, Miniprep)
31. Preparation of Probe
32. Isolation of High Molecular Weight Plant Genomic DNA and Mitochondrial (mt.) DNA
33. Restriction Fragment Length Polymorphism (RFLP)
34. Isolation of Molecular Markers Using Random Amplified Polymorphic DNA (RAPD)
35. SCARS and CAPS, PCR-Based Markers for Genetic Analysis
36. Site Directed Mutagenesis
37. Isolation of RNA and Northern Blotting
38. Isolation of Chloroplast DNA
39. DNA Sequencing
40. Identification of Proteins by Two-Dimensional Polyacrylamide Gel Electrophoresis (PAGE) - Based analysis and Western Blotting

APPENDICES
1. Components and their concentrations used in different plant tissue culture media
2. Commonly used plant growth regulators
3. Plant growth regulators, their preparation and storage
4. Conversion of mg/litre of μM concentrations for commonly used plant grwoth regulators
5. Common conversions of nucleic acids and proteins
6. List of equipments required for molecular biology laboratory