I am doing this experiment for my project. I didnt get results.I just want to know
You can follow the following link
http://genetics.wustl.edu/tslab/protocols/protein-stuff/iptg-induction/
Hallo Muthumani? For the IPTG induction, First, i hope your plasmids are ok with the necessary promoter. With respect to that, you need to check if your protein is toxic to the cells, check your protein with SDS PAGE and if the protein is well expressed but no activity/function, explore the possibility of inclusion bodies, expressing at different temperatures. All the best!!
What type of plasmid? What strain of bacteria? How did you check for induced protein?
for IPTG induction, first you check whether your plasmid have IPTG site for expression and then before and after induction with IPTG you have to collect cultures at different time intervals. whether the higher expression/OD was not observed means you have standardize the concentration of IPTG for induction. to high level of optimized OD values.
for IPTG induction, first you check whether your plasmid have IPTG site for expression and then before and after induction with IPTG you have to collect cultures at different time intervals. whether the higher expression/OD was not observed means you have to standardize the concentration of IPTG for induction. to get high level of optimized OD values.
Maybe you should say what procedure you followed, including cells, plasmid, IPTG concentration, gene of interest, temperature after induction, length of time after induction, optical density at the onset of induction.
When you say it didn't work, did you run a SDS-Page gel to check for induction or did you simply try an enzyme assay or purification?
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