Protocols in Biochemistry and Clinical Biochemistry offers clear, applied instruction to fundamental biochemistry methods and protocols, from buffer preparation to nucleic acid purification, protein, lipid, carbohydrate, and enzyme testing, and clinical testing of vitamins, glucose and cholesterol levels, among other diagnostics. Each protocol is illustrated with step-by-step instructions, labeled diagrams, and color images, as well as a thorough overview of materials and equipment, precursor techniques, safety considerations and standards, analysis and statistics, alternative methods and troubleshooting.
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Table of Contents
Unit I: Buffer, solution and pH1. Preparation of normal and molar solution2. Buffer preparation3. Determination of pKa, pH
Unit II: Nucleic Acids1. Isolation and Purification of DNA2. Isolation and Purification of RNA3. Purification of Nucleic Acid4. Quantitative estimation of Nucleic acid5. Agarose gel electrophoresis for separation of nucleic acid
Unit III: Carbohydrate and Lipids1. Qualitative test for carbohydrate (Benedict, Molisch etc)2. Qualitative test for Lipids3. Thin liquid chromatography (TLC for separation and identification of carbohydrate and lipids
Unit III: Proteins1. Isolation of Protein2. Qualitative test of Protein (Biuret test, Xanthoproteic test, Ninhydrin test etc)3. Estimation of Protein (Bradford, Lowry etc)4. Paper Chromatography for separation and identification of amino acid5. Ion exchange chromatography for separation of amino acid6. Protein purification7. SDS PAGE analysis of proteins8. Western blotting for protein analysis9. Isoelectric pH determination
Unit IV: Enzymes1. Enzyme kinetics2. Study of the activity of salivary amylase and the effect of pH, temperature, salts on its activity3. Enzymatic activity of Trypsin and Lipase4. Enzyme purification5. Determination of Km, Vmax and Ki
Unit V: Clinical test1. Acid and Alkaline phosphatase activity2. SGPT and SGOT test3. Estimation of Urea, Uric acid and Creatinine4. Estimation of Glucose5. Estimation of Cholesterol6. Estimation of Vitamins7. Glucose tolerance test8. Estimation of Ca2+ and T4 in serum9. Pregnancy test by HCG hormone10. Estimation of Glycosylated haemoglobin
Authors
Shweta Pandey Assistant Professor at the Department of Biotechnology, Govt VYT PG Autonomous College, Durg Chhatisgarh, India. Shweta Pandey is an Assistant Professor at the Department of Biotechnology, Govt VYT PG Autonomous College, Durg Chhatisgarh, India. Dr. Pandey received a PhD from the School of Life Sciences, Jawaharlal Nehru University (JNU) New Delhi, India. She has vast teaching experience and has published several papers in reputed international journals, many book chapters in edited books, and co-edited Protocols in Biochemistry and Clinical Biochemistry (Elsevier, 2020). Her research interests include cell cycle regulation, molecular and cell mechanisms of tumor cells, and cancer biology. Shyamal K Goswami Professor (retired), School of Life Sciences, Jawaharlal Nehru University, New Delhi, India. Shyamal Goswami has a research career spanning 35 years. He has experience teaching in biochemistry, cellular and molecular biology and redox biology at the postgraduate level. He served as Associate Professor and Professor in the School of Life Sciences at Jawaharlal Nehru University, India, from 1998 to 2020. Prior to this he was a Research Assistant Professor in the Department of Anatomy and Cell Biology at the State University of New York-Health Science Centre at Brooklyn, New York, USA.Dr. Goswami has contributed numerous publications across his field and has more than 1500 citations. He also co-authored Protocols in Biochemistry and Clinical Biochemistry and co-edited Post-transcriptional Gene Regulation in Human Disease, both published by Academic Press/Elsevier. His main areas of research are cell signaling and gene expression in the cardiovascular system; redox biology of degenerative diseases; and signal scaffolds and cell functions. Buddhi Prakash Jain Assistant Professor, Department of Zoology, School of Life Sciences, Mahatma Gandhi Central University Bihar, Motihari, India. 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.
