Workshop was started on 4th October, 2016 at 9.00 am. The participants (M.Sc. and B.Sc. (IADC) and M.Sc. (Mount Carmel)) were welcomed by Dr. Susanta Pahari. Followed by an hour of introductory lecture titled “Basics Of Protein Chromatography”, giving also overview of the workshop. The session was taken over by Mrs. Pushpa Reddy and Mrs Ramya Kumari B.S, giving the introduction to how to process the samples, introducing the technique of ammonium sulphate precipitation and calculations at every stage. Students started their hands on at 11.00 am, lab session ended by putting the sample for dialysis at 6.00 pm.
The session started by Ms Kokila, S. at 9.00 am by giving introduction to Ion exchange chromatography – why and when they are used during purification. The lab session was started by packing matrix into column, followed by wash and equilibration. The day ended by loading of protein sample onto column.
The session started by introduction to elution by Ms Kokila, S. The elution of enzyme was followed by screening of fractions for enzyme activity and protein concentration by UV-Vis spectroscopy. PAGE activity visualization of the crude, ammonium sulphate and the load was done. The session for the day ended with peak fractions pooling and dialysis.
The session started at 9.00am by Ms. Kokila, S. with the analysis of the final pool units. Gel visualization of the activity and the protein bands by silver staining were performed. Post lunch session was started by Mrs. Malathi with a lecture of “PAGE – basics and application”, followed by Mrs Pushpa Reddy and Mrs Ramya Kumari B.S explaining the participants about percentage yield, fold purification and their doubts in calculations. Dr. Susanta Pahari concluded the workshop with small discussion about the outcome of the workshop, followed by certificate distribution and taking feedback from participants. On the whole the participants were happy about the workshop and were willing to participate if conducted again.
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