Techniques in Molecular Biology I
Teacher
Dr. Satyabrata Nanda
Category
Core Courses
Course Attendees
Still no participant
Course Reviews
Still no reviews
Code (Credit): MMBB0504 (0-3-0)
Course Objectives
- To get a basic overview of molecular biology techniques, good lab practices and recombinant DNA technology.
- To get a hands on training in nucleic acid analysis, protein analysis, chromatography, and bacterial and phage genetics.
CO-PO Mapping
PSO1 : Impart knowledge and understanding of fundamental concepts and techniques of maintaining and enhancing soil fertility, crop production, crop management, crop improvement, biodiversity and sustainability of agriculture worldwide.
PSO2 : Develop the ability to know farming practices and their scope to improve in the rural areas and the environments.
PSO3 : Apply knowledge of basic science through Agricultural microbiology, Plant biochemistry and Biotechnology.
PSO4 : Apply knowledge of Agri-business management and finance to develop profitable agricultural system.
Syllabus
Practical
Good lab practices, preparation of buffers and reagents. Principle of centrifugation and spectrophotometry. Growth of bacterial culture and preparation of growth curve. Isolation of Genomic DNA from bacteria. Isolation of plasmid DNA from bacteria. Growth of lambda phage and isolation of phage DNA. Isolation of plant DNA (e.g. Rice/Moong/Mango/Merigold). Quantification of DNA by (a) Agarose Gel electrophoresis and (b) Spectrophotometry. PCR using isolated DNA. PAGE Gel electrophoresis. Restriction digestion of plasmid and phage DNA, ligation, Recombinant DNA construction. Transformation of E. coli and selection of transformants. Chromatographic techniques: (a) TLC, (b) Gel Filtration Chromatography, (c) Ion exchange Chromatography, (d) Affinity Chromatography. Dot blot analysis, Southern hybridization, and Northern hybridization. Western blotting and ELISA. Radiation safety and non-radio isotopic procedure.
References
Sambrook, J., and Russell, R.W (2001) Molecular cloning: A laboratory manual 3rd Edition, Cold spring harbor laboratory press, cold spring harbor, New York.
Wilson, K., and Walker, J., (2018) Principles and Techniques of Biochemistry and Molecular Biology 8th edition, Cambridge University Press.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A.,&Struhl, K., (2002) Short Protocols in Molecular Biology 5th edition, Current Protocols publication.
Session Plan
Session 1
Good lab practices.
Session 2-4
Preparation of buffers and reagents and their sterilization.
Session 5
Principle of centrifugation.
Session 6
Principle of spectrophotometry.
Session 7-10
Growth of bacterial culture and preparation of bacterial growth curve.
Session 11-12
Isolation of Genomic DNA from bacteria.
Session 13-14
Isolation of plasmid DNA from bacteria.
Session 15-16
Growth of lambda phage.
Session 17-18
Isolation of phage DNA.
Session 19-20
Isolation of plant DNA.
Session 21
Quantification of DNA by Agarose Gel electrophoresis.
Session 22
Quantification of DNA by Spectrophotometry.
Session 23-25
PCR using isolated DNA (RAPD/ISSR/SSR).
Session 26
PAGE Gel electrophoresis.
Session 27-28
Restriction digestion of plasmid.
Session 29-30
Restriction digestion of phage DNA.
Session 31-32
Ligation of DNA.
Session 33-34
Recombinant DNA construction.
Session 35-36
Transformation of E. coli.
Session 37-38
Selection of transformants.
Session 39
Chromatographic techniques - TLC.
Session 40
Chromatographic techniques - Gel Filtration Chromatography.
Session 41
Chromatographic techniques - Ion exchange.
Session 42
Chromatographic techniques - Affinity Chromatography.
Session 43
Dot blot analysis.
Session 44
Southern hybridization.
Session 45
Northern hybridization.
Session 46
Western blotting.
Session 47
ELISA.
Session 48
Radiation safety.
E-materials
E-materials have been embedded in the respective session plans. Click on the session plans to get the linked e-materials.
Question Bank
Assignment topics
Our Main Teachers
Dr. Satyabrata Nanda has a Ph.D. in Biotechnology with a specialization in Plant Protection. His education includes B-Tech, M-Tech, and Ph.D. in Biotechnology. He’s received the Overseas Postdoctoral Fellowship from the Chinese Academy of Agricultural Sciences for research at China National Rice Research Institute. His research is primarily focused on plant protection by using functional […]