Techniques and Protocols in Molecular Biology delivers detailed practical guidance for a variety of molecular techniques and procedures. Each chapter focuses on a different technique including isolation techniques for DNA and RNA, gene cloning, PCR, ELISA, and cell culture. These protocols include a detailed description of the technique along with the main principles, required equipment, and comprehensive steps to assist researchers with replicating the protocol and performing the techniques themselves.
This foundational reference is essential for students and researchers looking to enhance their understanding of key molecular techniques and reproduce them successfully in their own laboratory work.
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Table of Contents
1. Genomic DNA isolation (Bacteria, plant, and animal cells/tissues)
2. RNA isolation from mammalian cells/tissues, Drosophila, and Caenorhabditis elegans using monophasic lysis reagent
3. Reverse-transcription polymerase chain reaction
4. Quantitative reverse-transcription polymerase chain reaction: Real-time polymerase chain reaction
5. Agarose gel electrophoresis
6. 3’RACE (random amplification of cDNA end
7. 5’RACE (Random amplification of cDNA end
8. Plasmid isolation and purification (Alkaline lysis method)
9. Preparation of competent cells (Bacterial cells) and transformation (Hanahan’s method)
10. Molecular gene cloning
11. End-labeling of DNA (5’end labeling by T4 polynucleotide kinase)
12. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresis
13. Western blotting
14. Electrophoretic mobility shift assay for DNA-protein interaction
15. Silver staining of proteins in acrylamide gel compatible with mass spectrometry: Acidic method (From Shevchenko et al., 1996)
16. Animal cell culture (Adherent cells)
17. Cell viability by MTT assay
18. Lactate dehydrogenase assay
19. Luciferase assay
20. Co-immunoprecipitation
21. Protein purification (His-tagged proteins by immobilized metal affinity chromatography)
22. Subcellular fractionation by differential centrifugation (Separating nuclear, membrane, and cytosolic fraction)
23. DNase 1 footprinting
24. Chromatin immunoprecipitation-qPCR
25. Flow cytometry for cell surface markers
26. Basics of bioinformatics
27. GST pull down assay
28. Fluorescence In Situ hybridization from paraffin-embedded tissues
29. An overview of yeast two-hybrid assay
30. Enzyme-linked immunosorbent assay
