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. - Explores a wide range of advanced methods that can be applied by researchers in molecular biology and biotechnology - Features clear, step-by-step instruction for applying the techniques covered - Offers an introduction to laboratory protocols and recommendations for best practice when conducting experimental work, including standard operating procedures for key equipment

Dr. Khalid Zaffar Masoodi, a young scientist working as an Assistant Professor at Division of Plant Biotechnology in Sher-e Kashmir University of Agricultural Sciences and Technology of Kashmir (India) and holds Masters and Ph.D in Biotechnology. After completing his Ph.D., he worked in Prof. Zhou Wang's Lab at University of Pittsburgh, PA, USA as Postdoctoral Associate (2010-2014). Dr. Masoodi is a recipient of SERB Early Career Research award 2016, Travel Award, SBUR, USA 2012, Society of Endocrinology Journal Award 2018 and many more. With a glorious academic career, Dr. Masoodi holds a distinction in qualifying CSIR-UGC net in Life Sciences, DBT-BET JRF and has qualified Graduate aptitude test in Engineering and Sciences (GATE) while he was doing his Masters in Plant Biotechnology.

His lab (Transcriptomics Laboratory, also called as K-Lab inspired by Klenow fragment that revolutionized Biotechnology research) at SKUAST-K is well equipped with Fluorescence imaging facility, RT-PCR, cell culture, Western blotting and many state-of-the-art instruments. His research focus is on multi-dimensional work on Transcriptomics of stress biology (biotic/abiotic) in plants and Cancer biology. Apart from that, he is extensively working on drug discovery from medicinal plants. He has produced high-impact papers published in Oncogene, Molecular Cancer Therapeutics, Endocrinology, Journal of Functional Foods, Plos One, and many more, and has over 230 sequences in Genbank. He also deciphered the whole transcriptome of six Kashmiri apple varieties. Currently, he has three externally funded projects from SERB, DST which focus on high throughput screening of Medicinal plants for prostate cancer and cisgenic apple. In 2019-20 Dr. Masoodi filed 5 patent applications with the Indian Patent office.

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 DH5a 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