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.
1. Cover image - p. c 2. Title page - p. i 3. Copyright - p. iii 4. Table of Contents - p. v 5. Biography - p. xvii 6. Chapter 1 Genomic DNA isolation (Bacteria, plant, and animal cells/tissues) - p. 1 7. Chapter 2 RNA isolation from mammalian cells/tissues, Drosophila, and Caenorhabditis elegans using monophasic lysis reagent - p. 7 8. Chapter 3 Reverse-transcription polymerase chain reaction - p. 13 9. Chapter 4 Quantitative reverse-transcription polymerase chain reaction: Real-time polymerase chain reaction - p. 19 10. Chapter 5 Agarose gel electrophoresis - p. 27 11. Chapter 6 3′RACE (random amplification of cDNA end) - p. 31 12. Chapter 7 5′RACE (Random amplification of cDNA end) - p. 39 13. Chapter 8 Plasmid isolation and purification (alkaline lysis method) - p. 47 14. Chapter 9 Preparation of competent cells (Bacterial cells) and transformation (Hanahan's method) - p. 55 15. Chapter 10 Molecular gene cloning - p. 61 16. Chapter 11 End-labeling of DNA (5′end labeling by T4 polynucleotide kinase) - p. 67 17. Chapter 12 Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresis - p. 71 18. Chapter 13 Western blotting - p. 79 19. Chapter 14 Electrophoretic mobility shift assay for DNA-protein interaction - p. 89 20. Chapter 15 Silver staining of proteins in acrylamide gel compatible with mass spectrometry: Acidic method - p. 95 21. Chapter 16 Animal cell culture (Adherent cells) - p. 99 22. Chapter 17 Cell viability by MTT assay - p. 105 23. Chapter 18 Lactate dehydrogenase assay - p. 109 24. Chapter 19 Luciferase assay - p. 113 25. Chapter 20 Co-immunoprecipitation - p. 119 26. Chapter 21 Protein purification (His-tagged proteins by immobilized metal affinity chromatography) - p. 125 27. Chapter 22 Subcellular fractionation by differential centrifugation (Separating nuclear, membrane, and cytosolic fraction) - p. 131 28. Chapter 23 DNase 1 footprinting - p. 137 29. Chapter 24 Chromatin immunoprecipitation-qPCR - p. 143 30. Chapter 25 Flow cytometry for cell surface markers - p. 149 31. Chapter 26 Basics of bioinformatics - p. 153 32. Chapter 27 GST pull-down assay - p. 161 33. 27.1 Principle - p. 161 34. 27.2 Materials and equipment - p. 161 35. 27.3 Step-by-step method details - p. 162 36. 27.4 Expected outcomes - p. 164 37. 27.5 Advantages - p. 164 38. 27.6 Limitations - p. 165 39. 27.7 Optimization and troubleshooting - p. 165 40. 27.8 Safety considerations and standards - p. 165 41. 27.9 Alternative methods/procedures - p. 166 42. Chapter 28 Fluorescence In Situ hybridization from paraffin-embedded tissues - p. 167 43. 28.1 Principle - p. 167 44. 28.2 Materials and equipment - p. 167 45. 28.3 Step-by-step Method Details - p. 168 46. 28.4 Expected outcomes - p. 169 47. 28.5 Advantages - p. 170 48. 28.6 Limitations - p. 170 49. 28.7 Optimization and troubleshooting - p. 170 50. 28.8 Safety considerations and standards - p. 171 51. 28.9 Alternative Methods/procedures - p. 171 52. Chapter 29 An overview of yeast two-hybrid assay - p. 173 53. Chapter 30 Enzyme-linked immunosorbent assay - p. 177 54. References - p. 187 55. Index - p. 201
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.
1. Cover image - p. c 2. Title page - p. i 3. Copyright - p. iii 4. Table of Contents - p. v 5. Biography - p. xvii 6. Chapter 1 Genomic DNA isolation (Bacteria, plant, and animal cells/tissues) - p. 1 7. Chapter 2 RNA isolation from mammalian cells/tissues, Drosophila, and Caenorhabditis elegans using monophasic lysis reagent - p. 7 8. Chapter 3 Reverse-transcription polymerase chain reaction - p. 13 9. Chapter 4 Quantitative reverse-transcription polymerase chain reaction: Real-time polymerase chain reaction - p. 19 10. Chapter 5 Agarose gel electrophoresis - p. 27 11. Chapter 6 3′RACE (random amplification of cDNA end) - p. 31 12. Chapter 7 5′RACE (Random amplification of cDNA end) - p. 39 13. Chapter 8 Plasmid isolation and purification (alkaline lysis method) - p. 47 14. Chapter 9 Preparation of competent cells (Bacterial cells) and transformation (Hanahan's method) - p. 55 15. Chapter 10 Molecular gene cloning - p. 61 16. Chapter 11 End-labeling of DNA (5′end labeling by T4 polynucleotide kinase) - p. 67 17. Chapter 12 Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresis - p. 71 18. Chapter 13 Western blotting - p. 79 19. Chapter 14 Electrophoretic mobility shift assay for DNA-protein interaction - p. 89 20. Chapter 15 Silver staining of proteins in acrylamide gel compatible with mass spectrometry: Acidic method - p. 95 21. Chapter 16 Animal cell culture (Adherent cells) - p. 99 22. Chapter 17 Cell viability by MTT assay - p. 105 23. Chapter 18 Lactate dehydrogenase assay - p. 109 24. Chapter 19 Luciferase assay - p. 113 25. Chapter 20 Co-immunoprecipitation - p. 119 26. Chapter 21 Protein purification (His-tagged proteins by immobilized metal affinity chromatography) - p. 125 27. Chapter 22 Subcellular fractionation by differential centrifugation (Separating nuclear, membrane, and cytosolic fraction) - p. 131 28. Chapter 23 DNase 1 footprinting - p. 137 29. Chapter 24 Chromatin immunoprecipitation-qPCR - p. 143 30. Chapter 25 Flow cytometry for cell surface markers - p. 149 31. Chapter 26 Basics of bioinformatics - p. 153 32. Chapter 27 GST pull-down assay - p. 161 33. 27.1 Principle - p. 161 34. 27.2 Materials and equipment - p. 161 35. 27.3 Step-by-step method details - p. 162 36. 27.4 Expected outcomes - p. 164 37. 27.5 Advantages - p. 164 38. 27.6 Limitations - p. 165 39. 27.7 Optimization and troubleshooting - p. 165 40. 27.8 Safety considerations and standards - p. 165 41. 27.9 Alternative methods/procedures - p. 166 42. Chapter 28 Fluorescence In Situ hybridization from paraffin-embedded tissues - p. 167 43. 28.1 Principle - p. 167 44. 28.2 Materials and equipment - p. 167 45. 28.3 Step-by-step Method Details - p. 168 46. 28.4 Expected outcomes - p. 169 47. 28.5 Advantages - p. 170 48. 28.6 Limitations - p. 170 49. 28.7 Optimization and troubleshooting - p. 170 50. 28.8 Safety considerations and standards - p. 171 51. 28.9 Alternative Methods/procedures - p. 171 52. Chapter 29 An overview of yeast two-hybrid assay - p. 173 53. Chapter 30 Enzyme-linked immunosorbent assay - p. 177 54. References - p. 187 55. Index - p. 201
