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Laboratory III: Molecular Biology, Genetic Engineering and Bioinformatics (Practical) [1]

Paper Code: 
25MBL229
Credits: 
04
Contact Hours: 
120
Objective: 

The objectives of this course are to provide students with experimental knowledge of molecular biology and genetic engineering and bioinformatics.

 

 

Course Outcomes: 

 

Course

Learning outcome

(at course level)

Learning and teaching strategies

Assessment Strategies

Course Code

Course

Title

25MBL229

Laboratory III: Molecular Biology, Genetic Engineering and Bioinformatics

(Practical)

CO95: Estimation of quality and quantity of isolated plasmid and perform restriction digestion, and valediction of PCR, colony PCR.

CO96:Design experiments for gene transfer in prokaryotes and evaluate expression and regulation of gene and protein.

CO97:Applyfundamental bioinformatics tools and databases to analyze genetic sequences, interpret evolutionary relationships, and extract relevant biological insights from computational data.

CO98: Identify and illustrate the various elements ofmolecular biology, genetic engineering and bioinformatics

CO99: Discuss and defend the concepts of molecular biology, genetic engineering and bioinformatics and illustrate the exercises performed with appropriate methods and outcomes and maintain proper documentation of the same.

CO100: Contribute effectively in course-specific interaction

Approach in teaching:  Hands-on practical, demonstrations, simulations

 

 

Learning activities for the students:  Discussion,

Tutorials,

Assignments

Reading Journals

 

 

Class test, Semester end examination, Hands-on practical assessment
 

 

Experiments based on:

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

5. Plasmid DNA isolation and DNA quantitation

6. Restriction Enzyme digestion of plasmid DNA

7. Agarose gel electrophoresis

8. Polymerase Chain Reaction and analysis by agarose gel electrophoresis

9. Vector and Insert Ligation

10. Preparation of competent cells

11. Transformation of E.coli with standard plasmids, Calculation of transformation efficiency

12. Confirmation of the insert by Colony PCR and Restriction mapping

13. Expression of recombinant protein, concept of soluble proteins and inclusion body formation in E.coli, SDS-PAGE analysis

14. Purification of His-Tagged protein on Ni-NTA columns a) Random Primer labeling b) Southern hybridization

 

Bioinformatics

  1. Using NCBI and Uniprot web resources.
  2. Sequence information resource: Using NCBI, EMBL, Genbank, Swissprot/ TrEMBL, UniProt.
  3. Similarity searches using tools like BLAST and interpretation of results.
  4. Multiple sequence alignment using ClustalW.
  5. Use of gene prediction methods (GRAIL, Genscan, Glimmer).
  6. Use of different protein structure prediction databases (PDB).
  7. Construction and study of protein structures using Deepview/PyMol.
  8. Homology modelling of proteins.
  9. Use of miRNA prediction, designing and target prediction tools.

 

 

 

Academic Year: 
2025-2026 [2]

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Source URL: https://biotech.iisuniv.ac.in/courses/subjects/laboratory-iii-molecular-biology-genetic-engineering-and-bioinformatics-practical

Links:
[1] https://biotech.iisuniv.ac.in/courses/subjects/laboratory-iii-molecular-biology-genetic-engineering-and-bioinformatics-practical
[2] https://biotech.iisuniv.ac.in/academic-year/2025-2026