I have trouble to extract mRNA from macrophage / Candida coculture, I'm using 5mm acid washed beads in a tissue lyser, followed by classic RNA extraction (with columns). But I couldn't get any mRNA. Can someone recommend a better protocol or can advise me?
When I had to extract RNA from the top well of transwells, I used Trizol extraction.
I know it's an old-fashioned method, but it worked well as I could lyse the cells directly in the Trizol.
Dear Juliette,
We have tried several methods including bead beating, but found that the traditional grinding of N2 frozen cells in a mortar with pestle is the most efficient. Works better than enzymatic lysis of Candida. Alternatively, you might try some smaller beads, I believe we have used 0.5 or 0.4 mm acids washed glass beads.
Good luck
For physical grinding, use a bead dispenser.
Here is a bead dispenser that I work with. Also using a large number of samples. LabTIE also has a large variety of Bead dispensers, fitting tubes, vials, 96-well 384-well etc.
https://www.labtie.com/seed-bead-powder-dispensers/112346-bead-dispensers
https://www.labtie.com/seed-bead-powder-dispensers/112346-bead-dispensers
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