Hi,
I am trying to extract RNA from Pseudomonas aeruginosa biofilm adhered to falcon tube. Some protocols that said the biofilm should be washed with PBS and then resuspended in RNAlater. I have also read that EPS can interfere with RNA isolation and sonication is needed to release cells from biofilm.
Should I add RNAlater directly to washed biofilm and do the sonication process later, or should I resuspend the biofilm in PBS and do the sonication first, then centrifuge the tube and add RNAlater?
For side note, I'm using tiangen RNAprep pure ki for rna extraction.
Thanks in advance!
Add 1 mL of Trizol to the tube and agitate and sonicate. Then proceed with usual Trizol RNA isolation. You will experience too much RNA loss otherwise. The final RNA pellet bring up in 50 uL of nuclease-free water.
We work with H.influenzae. we scrap off the biofilm with PBS and put directly in RNA protect reagent from Qiagen.
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