MOLECULAR BIOLOGY
Course Length : Regular - 90 Days, Fastrack - 60 Days
Course Start : For Details Call +91 9247716307,
Limited seats per batch
Course Location : Hyderabad, India
Contact Us : Call +91 9247716307
 

Syllabus

 
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

 
 
 

 

Overview


 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

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