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…mehr
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.
Dr. Buddhi Prakash Jain is an Assistant Professor at the Department of Zoology, Mahatma Gandhi Central University, Motihari, Bihar, India. He completed his doctoral studies at the School of Life Sciences, Jawaharlal Nehru University (JNU) New Delhi, India. His research interests include gene expression regulation and endoplasmic reticulum stress in cancer. He is the recipient of several awards including a gold medal in Master Programme (M.Sc.), EMBO travel award, DST (Department of Science and Technology) Govt of India, Young Scientist travel award, and has published several papers in reputed international journals and a number of books and book chapters.
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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
