Hi, for my experiment I have to seed the cells on the lower (abluminal) side of the inserts (no cells on regular side). I am facing issues with isolating RNA from those cells. Anyone has any experience? There should be around 300,000 cells (on 0.4 um Falcon 12-well format inserts). I have tried Qiagen RNeasy columns for RNA isolation. Thanks.
Hi Anurag Paranjape
I am just asking out of curiosity!
Did you think of scrapping it from the bottom? Or use TRIzol and lyse the cells from the bottom?
Also, if you are not growing any cells inside the insert, why don't you flush it using TRIzol from inside the insert and collect the cells in a new 12 well culture dish?
Hi Ashis Kumar that's a great suggestion! Especially flushing out from top with trizol I am definitely going to try.. Thanks!
Hi Laurence Stuart Dawkins-Hall
thanks for those useful tips. I will be pooling 3 transwells for each condition, so hoping to get enough yield.- Badges
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