Techniques and Protocols in Molecular Biology

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.

• 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 reagentChapter 3 Reverse-transcription polymerase chain reactionChapter 4 Quantitative reverse-transcription polymerase chain reaction: Real-time polymerase chain reactionChapter 5 Agarose gel electrophoresisChapter 6 3’RACE (random amplification of cDNA endChapter 7 5’RACE (Random amplification of cDNA endChapter 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 cloningChapter 11 End-labeling of DNA (5’end labeling by T4 polynucleotide kinase)Chapter 12 Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresisChapter 13 Western blottingChapter 14 Electrophoretic mobility shift assay for DNA-protein interactionChapter 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 assayChapter 18 Lactate dehydrogenase assayChapter 19 Luciferase assayChapter 20 Co-immunoprecipitationChapter 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 footprintingChapter 24 Chromatin immunoprecipitation-qPCRChapter 25 Flow cytometry for cell surface markersChapter 26 Basics of bioinformaticsChapter 27 GST pull down assayChapter 28 Fluorescence In Situ hybridization from paraffin-embedded tissuesChapter 29 An overview of yeast two-hybrid assayChapter 30 Enzyme-linked immunosorbent assay