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