I isolated cells using magnetic beads, but I didn't remove the beads from the complex (sample-antibody-beads). They are still linked. However, I would like to know if is possible to extract the RNA from these cells while still coated with dynabeads (magnetic beads). Does someone have any clue/idea about the possibility to do this without compromise my sample?
This is possible. Mix the bead pellet (containing your cells) with a strong lysis reagent for RNA extraction - I would use Trizol or Nucleozol for this but I think the lysis reagents out of various RNA extraction kits would also work fine. Vortex and then ideally rotate the tubes end-over-end for a few minutes to make sure all the cells contact the lysis reagent and none are shielded by clumping of the beads. At this point all the RNA should be suspended in the lysis reagent and you can remove the beads (either by magnet or by centrifugation), transfer the supernatant into a fresh tube, and proceed with the RNA extraction. If you are wanting to use a column kit to purify the RNA, try to avoid putting beads into the column in case they block it up.
Thanks a lot Laura Leighton ! I really appreciated! You actually mentioned something that I have totally forgot before (about the use of column kit to purify the RNA). In fact I'm trying to avoid it from the beggining, but the principle that the beads could block the column can be applied for other similar/procedures as well... Thanks a lot for your answer and share your thoughts!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line