Laboratory II

Paper Code: 
MBL 229
Credits: 
7
Contact Hours: 
210
Objective: 

Course Outcomes (COs):

Course Outcomes

Learning and teaching strategies

Assessment Strategies

Upon completion of the course the learner will:

CO 90: Apply tools and techniques related to Bioinformatics

CO 91: Apply their knowledge to design immunological experiments

CO 92: Learn experiments related to Molecular Biology and Genetic Engineering 

CO 93: Design the experiments for studying dental carries susceptibility of individuals.

CO 94: Isolate and identify normal flora from different body regions.

CO 95:  Analyse the sensitivity of microbes against different antibiotics

Interactive Lectures,

Discussion,

Tutorials,

Demonstrations,

Assignments

Reading Journals

Class test, Semester end examinations, Quiz, Solving problems in tutorials, Assignments, Presentation, Individual and group projects

Immunology

  1. Selection of animals, preparation of antigens, immunization and methods of blood collection, serum separation and storage.
  2. Antibody titre by ELISA method.
  3. Double diffusion, Immuno-electrophoresis, Radial Immuno diffusion
  4. Complement fixation test
  5. Isolation and purification of IgG from serum or IgY from chicken egg.
  6. SDS-PAGE, Immunoblotting, Dot blot assays.
  7. Blood smear identification of leucocytes by Giemsa stain.
  8. Separation of leucocytes by dextran method.
  9. Demonstration of Phagocytosis of latex beads and their cryopreservation.
  10. Separation of mononuclear cells by Ficoll-Hypaque and their cryopreservation.
  11. Demonstration of ELISPOT.
  12. Demonstration of FACS.

 

Medical Microbiology

13.Preparation of blood smear and Enumeration of RBC and WBC

14. Isolation of Normal Flora from Skin

     15. Isolation and identification of Staphylococci and Streptococci from human body.

     16. Antibiotic sensitivity test by disc method

     17. Determination of MIC of antibiotics

     18. Normal Flora of Oral Cavity: Dental carries susceptibility test

 

 

 

 

Bioinformatics

  1. Using NCBI and Uniprot web resources.
  2. Introduction and use of various genome databases.
  3.  Sequence information resource: Using NCBI, EMBL, Genbank, Entrez Swissprot/ TrEMBL, UniProt.
  4. Similarity searches using tools like BLAST and interpretation of results.
  5.  Multiple sequence alignment using ClustalW.
  6.  Phylogenetic analysis of protein and nucleotide sequences.
  7.  Use of gene prediction methods (GRAIL, Genscan, Glimmer).
  8.  Using RNA structure prediction tools.
  9.  Use of various primer designing and restriction site prediction tools.
  10.  Use of different protein structure prediction databases (PDB, SCOP, CATH).
  11.  Construction and study of protein structures using Deepview/PyMol.
  12. Homology modelling of proteins.
  13.  Use of tools for mutation and analysis of the energy minimization of protein structures.
  14.  Use of miRNA prediction, designing and target prediction tools.

 

Molecular Biology and Genetic Engineering

  1.  Concept of lac-operon: a) Lactose induction of B-galactosidase. b) Glucose Repression. c) Diauxic growth curve of E. coli
  2. UV mutagenesis to isolate amino acid auxotroph
  3.  Phage titre with epsilon phage/M13
  4. Genetic Transfer-Conjugation, gene mapping Plasmid DNA isolation and DNA quantitation
  5. Restriction Enzyme digestion of plasmid DNA
  6. Agarose gel electrophoresis 8. Polymerase Chain Reaction and analysis by agarose gel electrophoresis
  7. Vector and Insert Ligation
  8. Preparation of competent cells
  9. Transformation of E. coli with standard plasmids, Calculation of transformation efficiency
  10. Confirmation of the insert by Colony PCR and Restriction mapping
  11. Expression of recombinant protein, concept of soluble proteins and inclusion body formation in E. coli, SDS-PAGE analysis
  12. Purification of His-Tagged protein on Ni-NTA columns a) Random Primer labeling b) Southern hybridization.

 

Academic Year: 
2023-2024
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