I'm working with E. coli for protein expression and I'd like to speed up the process if possible. The standard protocol is to add a 1:100 ratio for starter culture to the larger volume (ie. 10mL starter culture to a 1000mL flask). I'm wondering if it is possible to increase this to about 1:10 to increase the time it takes to get to .7 at OD600.
Thanks for your help in advance,
Daniel
I am questioning what is the aim of increasing the starting inocula. If you apply it just for obtaining cell mass probably it is OK, however, if there are many repeatedly experiments to be made under optimal conditions then you have to normalize the starting inocula during conducting all experiments
Dear Daniel
is it better to avoid this approach because if you are reducing the dilution factor from preculture to culture, you are speed up your culture but you also increase the amount of old cells and exausted media trasfered from pre-culture to culture and performance in terms of protein expression can be negativelly affected.
A possible solution, to obtain nice results but with more simple timelines is use an high cell density media instead lb (as the enpresso B media) that has longer exponential phase and you can induce also after o/n growth.
you can find more detail about it at page 2 of my blog: https://proteocool.blogspot.com/?m=1
good luck
Manuele
I am questioning what is the aim of increasing the starting inocula. If you apply it just for obtaining cell mass probably it is OK, however, if there are many repeatedly experiments to be made under optimal conditions then you have to normalize the starting inocula during conducting all experiments
- Badges
- Science method
- Similar topics
- Biological Science
- Genetics
- Gene Expression
- Protein Expression