The "Practical Handbook on Veterinary Biochemistry and Biotechnology" covers the analytical techniques commonly used in Veterinary Biochemistry and Biotechnology and can be used as a benchtop guide for undergraduate as well as post-graduate students of veterinary and animal sciences. The practical exercises covered in this book contain basic principles and working protocol for the techniques. The principal goal of the book is to provide students with full details of the practical steps necessary for carrying out each practical successfully by providing knowledge about the chemistry behind it. The book will enable the students and the researchers to develop a sound understanding of the principle and methodology of the biochemical and biotechnological techniques required to carry out basic and applied research in the subject. The topics have been selected following the VCI-MSVE syllabus, 2016 for Veterinary Biochemistry and some common Biotechnological techniques have also been included. In addition, descriptions about a few important Bioinformatics tools like sequence alignment and primer designing, etc. are also included to give an initial idea about these tools to the students. Finally, the authors hope and do believe that the book will be a great asset to the students to pursue their studies in Veterinary Biochemistry and Biotechnology.
Dr. Ankan De completed his M.V.Sc. and Ph.D. in Veterinary Biochemistry. He is working as Assistant Professor in the Department of Veterinary Physiology and Biochemistry, C.V.Sc. & A.H., R.K.Nagar, Tripura since 2011. He possesses over 10 years experience in teaching the subject of Veterinary Biochemistry and Biotechnology to Undergraduate students. He has implemented three major and two minor externally funded research projects. He has published more than 30 research articles in peer reviewed journals, 5 review/general articles and a book.
Dr. Jagan Mohanarao Gali, completed his Masters and Ph.D. from National Dairy Research Institute, Karnal and currently working as Assistant Professor in the Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University (Imphal), Aizawl, Mizoram. He possesses over 9 years experience in teaching the subject of Veterinary Biochemistry to Bachelors, Masters and doctoral students of Veterinary Science. He has published more than 50 research papers in international and national journals.
Dr. Parthasarathi Behera completed his M.V.Sc. from NDRI, Karnal and Ph.D. from IVRI, Izatnagar. He is working as Assistant Professor in the Department of Veterinary Physiology and Biochemistry, CVSc & AH, CAU(I), Aizawl since 2012. He is a core faculty member of BVSc & AH., MVSc and Ph.D. Dr. Behera has guided one MVSc student as major adviser and more than twenty students as member of advisory committee. Currently he is working as PI of a DST funded project. He has published more than 40 research articles in peerreviewed journals, 11 review/general articles and several book chapters.
Dr. Sani Nandi, M.V.Sc. (Veterinary Biochemistry), possesses more than 10 years work experiences which comprises both field oriented and academic activities. He has published 10 (Ten) numbers ofresearch papers, out of which 5 (Five) numbers of papers were published in different international journals. He had actively participated in multiple training programs, conferences and currently working as Assistant professor in the Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences and Animal Husbandry, R.K. Nagar, West Tripura.
1. Units of Measurement and Strength of Solution ......................... 1
2. Preparation and Standardization of Acids and Alkalies ........... 7
3. Preparation of Buffers and Determination of pH ..................... 13
4. Titration Curve of Acid Versus Base ........................................... 21
5. Physical Properties of Carbohydrates ......................................... 25
6. Reactions of Monosaccharides and Their Identification .......... 27
7. Reactions of Disaccharides and Their Identification ................ 41
8. Preparation of Osazone ................................................................. 45
9. Reactions of Polysaccharides and Their Identification ............ 49
10. Estimation of Lactose in Milk ........................................................ 53
11. Determination of Acid Number of Fat/Oil ................................ 55
12. Colour Reactions of Proteins ......................................................... 57
13. Precipitation Reactions of Proteins .............................................. 73
14. Identification of the Lipids............................................................. 77
15. Estimation of Amino Acids (Sorensen’s Method) ..................... 81
16. Effect of pH and Temperature on Enzyme Activity ................. 85
17. Collection of Blood and Separation of Plasma and Serum...... 89
18. Estimation of Blood Glucose .......................................................... 93
19. Estimation of Serum Total Proteins .............................................. 99
20. Electrophoretic Separation of Proteins ......................................105
21. Estimation of Serum Urea............................................................109
22. Estimation of Serum Cholesterol ................................................111
23. Estimation of Serum Bilirubin .....................................................115
24. Estimation of Serum Creatinine..................................................117
25 Estimation of Serum Uric Acid ...................................................119
26. Estimation of Electrolytes & Other Metabolic Intermediates of Blood.................121
27. Physical Examination of Urine ...................................................127
28. Microscopic Examination of Urine ............................................131
29. Normal Constituents of Urine.....................................................135
30. Abnormal (Pathological) Constituents of Urine ......................139
31. Isolation of Genomic DNA from Mammalian Cells ................143
32. Isolation of Genomic DNA from Bacteria .................................147
33. Isolation of Plasmid DNA from Bacterial Cells .......................153
34. Isolation of Total RNA from Mammalian Cells/Tissue .........157
35. Quantification and Purity Analysis of Nucleic Acids ............163
36. Polymerase Chain Reaction.........................................................167
37. Real Time PCR ...............................................................................171
38. Agarose Gel Electrophoresis ........................................................179
39. Directional Cloning of Target Gene(s) in Plasmid Vector(s).........................183
40. Extraction of Total Proteins Using SDS.....................................191
41. Analysis of Total Proteins by SDS-PAGE..................................193
42. Bioinformatics and the Databases ..............................................197
43. Sequence Alignment by Database Similarity Searching ........201
44. Primer Designing ...........................................................................207
45. ELISA: Principle, Protocol and Applications ...........................213
