RNA isolation from beads - (Apr/28/2008 )
Dear all, I want to capture something with antibody-coated dynabeads and isolate the RNA from them. I put trizol directly into the beads but I always have phenol contamination (peak on 270 nm, and ratio 260/230 of 0.25!). Is there anybody now how to deal with that?
Where's the phenol contamination come from actually? I assume that the beads degrade RNA somehow, but the nano drop still can read an amount of RNA but with low ratio 260/280 with the peak 270 nm. Does it mean that there's RNA there but with contamination?? If there's no RNA, can I still have phenol peak??
I rally appreciate any respons.
After adding Trizol, when you see the cells have been lyzed, you should put the tube on a magnet to remove the beads. If the sample is very viscous, this can take a while. Wait up to 2 min or dilute with a little more Trizol if needed. Transfer the lysate to a clean tube before continuing with your RNA isolation protocol.
Leaving the beads in lysis solution for too long may cause the surface of the beads to start to degrade and release iron ions which could influence your OD readings.