Advanced Methods in Molecular Biology and Biotechnology
A Practical Lab Manual
- 1st Edition - October 28, 2020
- Imprint: Academic Press
- Authors: Khalid Z. Masoodi, Sameena Maqbool Lone, Rovidha Saba Rasool
- Language: English
- Paperback ISBN:9 7 8 - 0 - 1 2 - 8 2 4 4 4 9 - 4
- eBook ISBN:9 7 8 - 0 - 1 2 - 8 2 4 4 5 0 - 0
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 biote… Read more
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Request a sales quoteAdvanced 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
Undergraduate and graduate students of molecular biology and / or biotechnology
- Cover image
- Title page
- Table of Contents
- Copyright
- Dedication
- Preface
- Foreword
- Acknowledgment
- Introduction
- About the author
- Disclaimer
- Author contribution
- Chapter 1: Introduction to molecular biology techniques
- Abstract
- Safety measures
- Preparation of the solutions
- Working with the nucleic acids
- Sterile techniques
- Chapter 2: Agarose gel electrophoresis
- Abstract
- Definition
- Rationale
- Before you begin
- Key resources table
- Materials and equipments
- Step-by-step method details
- Expected outcomes
- Quantification and statistical analysis
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 3: SDS-polyacrylamide gel electrophoresis
- Abstract
- Definition
- Rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Quantification and statistical analysis
- Advantages
- Limitations
- Optimization and troubleshooting
- Chapter 4: Immunoelectrophoresis
- Abstract
- Definition
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Chapter 5: Plasmid DNA isolation using alkaline lysis method from E. coli DH5α cells
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 6: Restriction digestion of plasmid DNA
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Chapter 7: Genomic DNA extraction from the plant leaves using the CTAB method
- Abstract
- Definition
- Rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Quantification and statistical analysis
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 8: Genomic DNA extraction from silkworm using modified phenol:chloroform method (Suzuki et al., 1972)
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 9: Genomic DNA extraction from Fungal Mycelium using the modified Cenis, 1992 method
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 10: Bacterial transformation using CaCl2
- Abstract
- Definition
- Rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 11: Transformation of E. coli by electroporation
- Abstract
- Definition
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 12: Transfection of cells using lipofectamine
- Abstract
- Definition
- Key resources table
- Materials and equipments
- Step-by-step method details
- Advantages
- Optimization and troubleshooting
- Chapter 13: Total RNA isolation
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 14: RNA isolation from plant tissues with high phenolic compounds and polysaccharides
- Abstract
- Definition and rationale
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Quantification and statistical analysis
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Chapter 15: Total protein extraction with TCA-acetone and 2D gel electrophoresis
- Abstract
- Definition
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected results
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Chapter 16: Dansyl-Edman method of peptide sequencing
- Abstract
- Definition
- Key resources table
- Materials and equipment
- Step-by-step method details
- Advantages
- Limitations
- Chapter 17: ISSR analysis
- Abstract
- Definition
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 18: Gene cloning/recombinant DNA technology
- Abstract
- Definition
- Rationale
- Materials and equipments
- Attributes of an ideal vector
- Attributes of a good host cell
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Chapter 19: Polymerase chain reaction (PCR)
- Abstract
- Definition
- Rationale
- Types of PCR
- Before you begin
- Key resources table
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Applications of PCR
- Applications of PCR in agricultural sciences
- Optimization and troubleshooting
- Alternative methods/procedures
- Chapter 20: Blotting/hybridization and its types
- Abstract
- Definition
- Types of blotting
- Step-by-step method details
- Step-by-step method details
- Step-by-step method details
- Step-by-step method details
- Chapter 21: Dot blot analysis
- Abstract
- Definition and rationale
- Before you begin
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Chapter 22: ELISA (enzyme-linked immunosorbent assay)
- Abstract
- Definition
- Rationale
- Materials and equipment
- Step-by-step method details
- Quantification and statistical analysis
- Advantages
- Limitations
- Chapter 23: Study of principle of centrifugation
- Abstract
- Definition
- Types of centrifuge
- Before you begin
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Limitations
- Optimization and troubleshooting
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 24: Study of principle of chromatography
- Abstract
- Definition
- Expected outcomes
- Advantages
- Limitations
- Chapter 25: Thin layer chromatography (TLC)
- Abstract
- Definition
- Rationale
- Materials and equipment
- Step-by-step method details
- Quantification and statistical analysis
- Advantages
- Limitations
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 26: Gel-filtration or size-exclusion chromatography
- Abstract
- Definition
- Rationale
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Alternative methods/procedures
- Chapter 27: Ion-exchange chromatography
- Abstract
- Definition
- Rationale
- Before you begin
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Advantages
- Alternative methods/procedures
- Chapter 28: To perform affinity chromatography
- Abstract
- Definition and rationale
- Key resources table
- Materials and equipment
- Step-by-step method details
- Advantages
- Limitations
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 29: To perform paper chromatography
- Abstract
- Definition and rationale
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Quantification and statistical analysis
- Advantages
- Limitations
- Safety considerations and standards
- Alternative methods/procedures
- Chapter 30: Growth of bacterial cultures and preparation of growth curve
- Abstract
- Definition
- Rationale
- Materials and equipment
- Step-by-step method details
- Expected outcomes
- Chapter 31: To study standard operating procedures (SOPs) for various equipment
- Abstract
- Autoclave operation
- Laboratory centrifuge
- Distillation unit
- Electrophoresis equipment
- Fridge and freezer
- Operating procedure
- General procedures
- Maintenance
- Gel documentation system
- Incubator/shaker
- Thermocycler/PCR
- General points
- Pipette
- Water bath
- Index
- Edition: 1
- Published: October 28, 2020
- Imprint: Academic Press
- No. of pages: 218
- Language: English
- Paperback ISBN: 9780128244494
- eBook ISBN: 9780128244500
KM
Khalid Z. Masoodi
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).
Affiliations and expertise
Assistant Professor/Junior Scientist at the Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, IndiaSL
Sameena Maqbool Lone
Sameena Maqbool Lone is a PhD scholar in the Division of Vegetable Science, Sher-e-Kashmir University of Agricultural Sciences and Technology, India. She holds a Master's degree in Horticulture from SKUAST-K, Shalimar. At the Master's level, she worked on "Heterosis and Combining Ability Studies in Cherry Tomato" and obtained competence in morphological and molecular characterization, as well as numerous plant tissue culture technologies. In her Phd programme, she is working on the “Morpho-Molecular Characterization and Nutraceutical Profiling of different carrot genotypes (Daucus carota L.)”. She has outstanding academic and extracurricular achievements, for which she received a merit scholarship from SKUAST-K during her Bachelor's and Master's degree programmes. She has contributed to over 23 academic papers, 6 books, and more than 100 new gene sequences in Genbank by 2022.
Affiliations and expertise
Division of Vegetable Science, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, IndiaRR
Rovidha Saba Rasool
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.
Affiliations and expertise
Division of Plant Pathology, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, IndiaRead Advanced Methods in Molecular Biology and Biotechnology on ScienceDirect