Team:METU Turkey/Results Discussion/QS results

 

Purification with QS BL21 pTriEx
This was the first experiment of BL21 pTriEx with FPLC. We obeyed the general protein purification procedure except sonication. We lysed pellets with French Press. Then we centrifued the crude lysate at 32000 g for 60 min. We took a sample for SDS-PAGE. Then we continue with filtration. Filtered crude lysate was looaded to Q-sepharose column. According to FPLC graph, we have a sharp peak at 280nm which represents proteins. We picked up the tubes between 9 and 14. SDS-Page shows the sharp peak that contains CooA. F11 is the top of the sharp peak. We also check small peak which corresponds to 46th tube. We combined the F10, F11 and F12 elutes for further purification with chelating sepharose column.