Molecular Biology and Recombinant DNA Technology : Practical Manual Series Vol. II

Contents: Preface. 1. Introduction and history of molecular biology. 2. About gene. 3. Record making and keeping. 4. Safety guidelines. 5. Safe handling of microorganisms. 6. Preparation of buffers and solutions. 7. Purification of the Plasmid DNA. 8. Standard curve for molecular mass of protein. 9. DNA extraction from Agarose Gel. 10. Melting and annealing temperature. 11. DNA sequencing by ABI automated DNA sequencer. 12. SDS PAGE an introduction. Exercises: 1. Aseptic technique and establishing pure cultures. 2.  Preparation of glycerol stocks. 3. Preparations of culture media. 4. Growth in liquid medium. 5. Determination of viable cells. 6. Determination of cell mass. 7. Determination of cell number. 8. Growth curve. 9. Use of micropipettes. 10. To study multiple Allele in human blood. 11. Isolation of DNA from animal tissue blood. 12. Isolation of genomic DNA from blood 2. 13. Isolation of DNA from plant leaves C TAB method. 14. Isolation of high molecular weight genomic DNA from bacteria. 16. Isolation of plasmid DNA from E. coli bacteria. 17. Plasmid isolation Alkaline lysis method. 18. Large scale isolation of Plasmid DNA by Column chromatography. 19. Preparation of DNA from cell line high salt method. 20. DNA preparation form adherent cells. 21. Purification of DNA. 22. Concentration of DNA. 23. Quantitative estimation of DNA by spectrophotometric method. 24. Production of cDNA. 25. Gel electrophoresis. 26. Pulse field gel electrophoresis. 27. RAPD analysis for genetic similarity. 28. RNA isolation from tissue culture cells. 29. Polymerase chain reaction (PCR). 30. Preparations purification and hybridization of probe. 31. Preparation of high titer from bacteriophage. 32. Preparation of competent cells. 33. Transformation. 34. Bacterial transformation using electroporation. 35. Screening bacterial colony by hybridization. 36. Transformation of saccharomyces cereviciae. 37. To perform restriction digestion of genomic DNA/plasmid DNA by E. co RI. 38. Restriction digestion with the enzymes Hind III sal I.  39. Estimeation of proteins by Lowrys method. 40. Estimation of proteins by Bradfords method. 41. To perform SDS polyacrylamide  gel electrophoresis of proteins. 42. Determination of Amino Acid composition. 43. Southern blotting. 44. Northern blotting. 45. Western blotting. 46. Qualitative assay for B Galactosidase in  yeast colonies.  47. Determination of B. galactosidase in permeabilized yeast cells. 48. Assay of B galactosidase in cell extracts. 49. Immunoscreening of Lambda gt II bacteriophage libraries with antibody probes. 50. Isolation of T cell from synovial membranes. Bibliography. Appendices.