Low yield at His tagged-peptide production in labeling media - (Jan/22/2006 )
Hi all,
I have been trying to produce 86 aa Histagged peptide in both LB and labeling (ennriched with C13 and N15) media. For LB, my yield is about 7-8 mg per liter which is reasonable. But the labeling media could not work so well. I made time course study for IPTG induction and noticed that the peptide started to be degraded after induction in labeling media. Also, I decrased the incubation temperature to 28C. Without using IPTG, I incubate 50ml culture for 8h and took samples at each hour and checked the expression by SDS-PAGE, it seems that fourth hour is the highest but still I have low yield problem when I work with large scale.
Is there anyone that has worked with labeling media and met any yield problem?
I appriciate any idea..
Thanks..
I have been trying to produce 86 aa Histagged peptide in both LB and labeling (ennriched with C13 and N15) media. For LB, my yield is about 7-8 mg per liter which is reasonable. But the labeling media could not work so well. I made time course study for IPTG induction and noticed that the peptide started to be degraded after induction in labeling media. Also, I decrased the incubation temperature to 28C. Without using IPTG, I incubate 50ml culture for 8h and took samples at each hour and checked the expression by SDS-PAGE, it seems that fourth hour is the highest but still I have low yield problem when I work with large scale.
Is there anyone that has worked with labeling media and met any yield problem?
I appriciate any idea..
Thanks..
at first growing cell in LB until the OD is 0.6,then collect the culture and transfer it to the M9 media(ennriched with C13 and N15),growing cell for another 1h,then induction wity iptg.
very time the yield is very good
good luck