I could like to introduce myself as gokila doing doctorate in proteomics lab . just a few months back ,i have joined the lab and my starting work is to overexpress the recombinant protein which is in BL-21 strain. This overexpression was already done by two people ,now i have been contuining it.., THE FIRST PERSON CLONED AND OVEREXPRESSED THE PROTEIN AT CONC. 60 MICROMOLAR IPTG .THE SECOND PERSON CARRIED OUT THE WORK AFTER 1 YEAR - OVEREXPRESSED AND PURIFIED THE PROTEIN AT CONC. 0.1 MILLIMOLAR IPTG . I HAVE TRYED BOTH THE CONDITIONS WHAT THEY HAVE MENTIONED IN THEIN REPORTS - 3 HOURS GROWTHING HTE CELLS AND INDUCED IN THE OD 0.7 TO 0.8 AND INDUCED LEFT FOR 4 HOURS AND ANALYSED. BUT ,I AM NOT GETTING THE INDUCED BAND AT 62KDA .
Please help me to get start with my doctorate work ..,