Day 1: Monday 23 September

8:45-9:00         Welcome/ Meet and Greet

9:00-9:15         Workshop Overview

9:15-9:45         Overview of recombinant protein expression

9:45-10:15       Lab activity: Launch culture of bacteria cells containing fusion plasmid

10:15-10:30     Coffee break

10:30-11:30     Principles of cloning, cloning strategies and cloning vectors

11:30-12:30     Selection of recombinants, protein expression and purification

12:30-13:30     Lunch

13:30-13:45     Lab activity: Measurement of optical density of bacteria culture

13:45-14:00     Lab activity: Induction of bacteria culture

14:00-14:45     Construction of cDNA library and selection of specific clone from cDNA library

14:45-15:00     Break

15:00-16:00     Construction of chimeric antigens and codon optimization

16:00-17:00     Lab activity: Harvesting of bacteria cells and resuspension in lysis buffer

 

Day 2: Tuesday 24 September

9:00-9:15         Lab activity: Lysis of bacteria cells

9:15-9:45         Lab activity: Collection of lysate supernatant by centrifugation

9:45-11:15       Lab activity: Incubation of resin and supernatant

11:15-12:15     Lab activity: Purification using amylose resin

12:15-13:45     Lab activity: Incubation of purified protein with nickel resin

12:15-13:15     Lunch

13:15-13:45     Western blot overview

13:45-15:15     Lab activity: Purification using nickel resin

15:15-16:15     Lab activity: SDS PAGE

15:30-16:00     Break

16:15-16:45     Lab activity: Staining of gel

16:30-17:00     Lab activity: Transfer of protein to nitrocellulose membrane overnight

16:45-17:00     Lab activity: Destaining of gel overnight

 

Day 3: Wednesday 25 September

9:00-9:15         Lab activity: Viewing of destained gel

09:15-11:15     Lab activity: Blocking of membranes with milk and continuation Western blot procedures

10:00-10:30     Coffee break

10:30-11:15     Application of recombinant proteins

11:15-13:30     Lab activity: Washing and blocking of nitrocellulose membrane

12:00-13:00     Lunch

13:30-15:30     Lab activity: Incubation with secondary antibody

15:30-16:00     Lab activity: Washing and revealing of nitrocellulose membrane

16:00-17:00     Q&A


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