Projects and Trainings offered in Molecular Biology Department
Module I: Nucleic acids
Introduction to nucleic acids structure and properties
DNA isolation from
Biological fluids
Animal tissues
Plants
Bacteria
Plasmid isolation
Purification of nucleic acids
Ethanol Precipitation
Salt precipitation
Gel elution by phenol method
Gel elution by glass wool method
Characterization of nucleic acids
Integrity
DNA staining
Quantification
Estimation by diphenylamine method
Spectrophotometric method
Kinetics
Denaturation kinetics (Tm)
Renaturation kinetics
RNA isolation from
Biological fluids
Animal tissues
Quantification of RNA by spectroscopy
Restriction analysis
Restriction mapping (RFLP)
Plasmid foot printing
Module II: Proteins
Extraction and characterization of proteins
Cell lysis methods
Isolation of proteins from culture supernatants
TCA Precipitation
Acetone Precipitation
Colorimetric Quantification of proteins
Biuret method
Lowry method
Bradford
Absorption at A280 A205
Enzymology
Screening for enzyme producing organisms
Isolation of enzymes from different bacterial cultures
Purification of enzymes
Mutation studies
Enzyme Kinetics
Effect of temperature
Effect of pH
Effect of substrate concentration
Enzyme assays
Module III: Polymerase Chain Reaction (PCR)
Designing Primers for candidate genes
Standardization of primer conditions
MgCl2concentration
Annealing temperature
Annealing time
DNTPs/MgCl2 ratio
Elimination of PCR based errors by using different polymerases
Conventional PCR
Hot start PCR
Allele Specific PCR
Multiplex PCR
Semi quantitative PCR
Reverse Transcriptase PCR
Real time PCR (Demo)
Site Directed Mutagenesis
Creation of mutants by incorporation of mutant base in primer
Mega-primer based site directed mutagenesis
Site directed mutagenesis by whole plasmid amplification
Mutation Detection Methods
Restriction fragment length polymorphism
Hetero-duplex analysis
Single Strand Conformation Polymorphism
Module IV: Recombinant DNA technology
Culturing microorganisms
Selection of medium
Antibiotic Selection
Mini-prep preparation
Creation of sticky ends by Restriction endonuclease digestion
Creation of Blunt ends by Restriction endonuclease digestion
Amplification of fragments containing T overhangs
Ligating into cloning and expression vectors
Ligation of sticky end fragments
Ligation of blunt end fragments
T/A ligation
Enzyme free cloning
Preparation of competent host cells
Preparation of ultra competent cells
Introducing hybrid DNA in to host cells by transformation
Screening positive clones
Plasmid size analysis
Colony PCR
Releasing insert with restriction enzymes
Overexpression of the cloned fragments by induction
Isolation and purification of overexpressed proteins
