Advanced Methods in Molecular Biology and Biotechnology: A Practical Lab Manual is a concise reference on common protocols and techniques for advanced molecular biology and biotechnology experimentation. Each chapter focuses on a different method, providing an overview before delving deeper into the procedure in a step-by-step approach. Techniques covered include genomic DNA extraction using cetyl trimethylammonium bromide (CTAB) and chloroform extraction, chromatographic techniques, ELISA, hybridization, gel electrophoresis, dot blot analysis and methods for studying polymerase chain reactions. Laboratory protocols and standard operating procedures for key equipment are also discussed, providing an instructive overview for lab work.
This practical guide focuses on the latest advances and innovations in methods for molecular biology and biotechnology investigation, helping researchers and practitioners enhance and advance their own methodologies and take their work to the next level.
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1. Introduction to molecular biology techniques 2. Agarose gel electrophoresis 3. SDS-polyacrylamide gel electrophoresis 4. Immunoelectrophoresis 5. DNA plasmid isolation using Alkaline Lysis Method from E. coli DH5? cells 6. Restriction digestion of Plasmid DNA 7. Genomic DNA extraction from plant leaves using CTAB method 8. Genomic DNA extraction from silkworm using modified phenol: chloroform method (Suzuki et al., 1972) 9. Genomic DNA extraction from Fungal Mycelium using modified Cenis, 1992 method 10. Bacterial transformation protocol using CaCl2
11. Transformation of E. coli by electroporation 12. Transfection of cells using lipofectamine 13. Total RNA isolation 14. RNA isolation from plant tissues with high phenolic compounds and polysaccharides 15. Total protein extraction with TCA-acetone and 2D gel electrophoresis 16. Dansyl-Edman method of peptide sequencing 17. ISSR analysis 18. Gene cloning/recombinant DNA technology 19. Polymerase Chain Reaction (PCR) 20. Blotting/ hybridization and its types 21. Dot blot analysis 22. ELISA (enzyme-linked immunosorbent assay) 23. Study of principle of centrifugation
24. Study of principle of chromatography
25. Thin layer chromatography (TLC)
26. Gel-filtration or size-exclusion chromatography
27. Ion-exchange chromatography
28. To perform affinity chromatography
29. To perform paper chromatography
30. Growth of bacterial cultures and preparation of growth curve
31. To study standard operating procedures (SOPs) for various equipment
Dr. Khalid Zaffar Masoodi is a young scientist who works as an Assistant Professor in the Division of Plant Biotechnology at the Sher-e Kashmir University of Agricultural Sciences and Technology in Kashmir (India). He holds a Master's degree in Biotechnology and a doctorate in Plant Biotechnology. Dr. Masoodi is the recipient of numerous awards, including the SERB Early Career Research Award 2016, the SBUR Travel Award in the United States of America in 2012, and the Society of Endocrinology Journal Award in 2018 at Glasgow United Kingdoms. Dr. Masoodi has a distinguished academic record, having qualified for the CSIR-UGC net in Life Sciences, the DBT-BET JRF, and the Graduate Aptitude Test in Engineering and Sciences (GATE) while doing his Masters in Biotechnology. Dr. Masoodi has filed Six patent applications in last 2 years, has over 12 years of teaching and research experience, and has published numerous scientific articles in the fields of cancer research, human disease research, and biotechnology research. His laboratory (Transcriptomics Laboratory, often known as K-Lab, after the Klenow fragment) at SKUAST-K is fully equipped with fluorescence imaging, real-time PCR, cell culture, and Western blotting including other state-of-the-art equipment. His research interests span multiple dimensions, including transcriptomics of plant stress biology (biotic/abiotic) and cancer biology. Apart from that, he is extensively involved in the research of new drugs from medicinal plants. Additionally, he is a co-author of Advanced Molecular Biology and Biotechnology Methods (Academic Press, 2020).
Sameena Maqbool Lone Division of Vegetable Science, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, India.
Sameena Maqbool Lone, currently, an M.Sc. scholar in the Division of Vegetable Science, SKUAST-K, did her Bachelor's in Horticulture, from SKUAST-K, Shalimar. She is working on the Heterosis and Combining Ability Studies in Cherry tomato (Solanum spp.) and various technologies related to plant tissue culture under the co - guidance of Dr. Khalid Z. Masoodi and is also holding an excellent academic and extracurricular record. She was also awarded merit scholarship by SKUAST-K during her Bachelor's degree programme.
Rovidha Saba Rasool Division of Plant Pathology, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, India.
Rovidha Saba Rasool did her Bachelors in Agriculture from SKUAST-K, Wadura followed by Masters in Plant Pathology from SKUAST-Kashmir, Shalimar. Currently, she is pursuing her Ph.D from Division of Plant Pathology, SKUAST-Kashmir. She has done commendable work in finding new genes involved in the pathogenesis of shot hole in stone fruits through insertional mutagenesis in Thyrostroma carpophilum. She has been a good student holding an excellent academic and extracurricular record and was awarded Ph.D. merit scholarship by SKUAST-K, in 2018-19. Apart from this, Rovidha has taken part in various national-level sports competitions and has won many national and state-level awards in baseball and handball.