I am trying to extract RNA from Escherichia coli grown in M9 minimal media supplemented with different sugars, however, every time the RNA extract ranges from 25 to 50 ng/ul.
I am using the Qiagen Rneasy mini kit. Do you have any idea how to increase the yield? Or can i go ahead with this low yield for further downstream applications?
If i am going to do a Dnase treatment, i am going to use all RNA extract for Dnase treatment of 1 ug RNA. Is that ok?
Looking forward for your help and advice.
Dear Nehal Aasem
You have to increase the number of E.coli cells up to 1010 cells /ml and proceed extraction
I would ask are your cultures growing to appropriate density? Do you get different results with cells grown in LB? you need to determine whether the problem is growth or your RNA extraction protocol.
@aly savadogo I increased but i was afraid that a great increase would block the column in the Rneasy kit
@David i am using K 12 so i checked them added Thiamine. I am extracting the RNA to check gene expression of certain sugar utilizing genes. Thank you for the note on catabolite repression
@Michael J.Benedik yes they are growing to a different density so the media is working fine , however the RNA yeild is always extremly low below 100 ng/ul
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