Techniques and Protocols in Molecular Biology

<p>1. Genomic DNA isolation (Bacteria, plant, and animal cells/tissues)<br>2. RNA isolation from mammalian cells/tissues, Drosophila, and Caenorhabditis elegans using monophasic lysis reagent<br>3. Reverse-transcription polymerase chain reaction<br>4. Quantitative reverse-transcription polymerase chain reaction: Real-time polymerase chain reaction<br>5. Agarose gel electrophoresis<br>6. 3&rsquo;RACE (random amplification of cDNA end<br>7. 5&rsquo;RACE (Random amplification of cDNA end<br>8. Plasmid isolation and purification (Alkaline lysis method)<br>9. Preparation of competent cells (Bacterial cells) and transformation (Hanahan&rsquo;s method)<br>10. Molecular gene cloning<br>11. End-labeling of DNA (5&rsquo;end labeling by T4 polynucleotide kinase)<br>12. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresis<br>13. Western blotting<br>14. Electrophoretic mobility shift assay for DNA-protein interaction<br>15. Silver staining of proteins in acrylamide gel compatible with mass spectrometry: Acidic method (From Shevchenko et al., 1996)<br>16. Animal cell culture (Adherent cells)<br>17. Cell viability by MTT assay<br>18. Lactate dehydrogenase assay<br>19. Luciferase assay<br>20. Co-immunoprecipitation<br>21. Protein purification (His-tagged proteins by immobilized metal affinity chromatography)<br>22. Subcellular fractionation by differential centrifugation (Separating nuclear, membrane, and cytosolic fraction)<br>23. DNase 1 footprinting<br>24. Chromatin immunoprecipitation-qPCR<br>25. Flow cytometry for cell surface markers<br>26. Basics of bioinformatics<br>27. GST pull down assay<br>28. Fluorescence In Situ hybridization from paraffin-embedded tissues<br>29. An overview of yeast two-hybrid assay<br>30. Enzyme-linked immunosorbent assay</p>