I have been growing cells on matrigel and subsequently lysing (commercial lysis buffer) in an attempt to isolate RNA. Is there any reason to think that the matrigel might be interfering with lysis? Is there a specific lysis protocol for cells grown on matrigel?
Have you find an explanation along this time? Because recently I extracted RNA from astrocyte culture on matrigel and the amount it was very small and I was not sure that it was correlated to the number of cells.
I haven't found an answer, but we did grow the cells on matrigel and lyse with Trizol, and we got enough RNA of high quality to make RNA-seq libraries. However, I don't have a control population of cells not grown on matrigel, so there's no way to tell whether that influenced the isolation or not.
Thank you.... I have even tried Trizol with cells on matrigel. I obtained RNA but I have to increase the number of cells because probably there is an effect about the amount that you can extract. Probably there is a better protocol but I don't know until now.
Hi guys, thanks for the answers.
so this means we can use Trizol to isolated RNA directly from cells which grow on matrigel?
can i do this?
I have not done this protocol for long time. I remember that sometime I collected good amount of RNA sometimes very low, and I didn't understand why. Now in my new laboratory we use RNA kit extraction from Invitrogen and I am so happy about performance: always high amount also from low number of cells, moroever you don't use anymore trizol that is very toxic.
Thanks for your point of view but if you want to sell your product this is not the appropriate place....let's stay consistent with the question (now it is also old; the conversation started in 2014)
Thanks Clara for your personal opinion!
Just to mention, some other researchers already highly appreciated my answer.
And no, its still not a product, its a project.
And no, although the question is from 2014, it is still a very actual one.
All the best for your reseach, Johannes
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