Hi to all.
I have some problems in extracting RNA from Ostreococcus tauri through the Trisure protocol. In this protocol, I collect the pellet of 25 ml from Ostreococcus culture by centrifuging during 4 min at 3200 g. Then, I wash the pellet with PBS 1X and centrifuge again. After this, I freeze the pellet in liquid nitrogen.
When I try to resuspend the pellet in Trisure (first step of Trisure protocol), it's impossible to resuspend it completely.
I have tried to grind the cells mechanically with 5 mm steal beads but it has not greatly improved the RNA extraction.
Can someone help me with this?
I believe that for RNA extraction RNAzol is recommended, though Trisure should work well. In RNAzol protocol you have to start with 400 microL of liquid, therefore you can use this volumen to resuspend the cells, instead of using directly the RNAzol (or Trisure) to resuspend the cells (maybe with Trisure you clog the pellet). This maybe improve resuspension/lysis?
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