I have tried isolating RNA from dead roots with no success. We call them dead just because they are discolored and black/dark brown but not sure if that means they are completely dead. I have tried many RNA isolation methods but none works for these particular roots.
Any insights?
Well, roots can be a bit tricky even when the tissue is fresh and healthy. It can help to really grind the samples well. I used a ceramic mortar and pestle for Arabidopsis roots and that wasn't easy. For tougher samples it can help to add a bit of sand (the initial centrifugation step removes the sand with other debris). Are you trying roots from soil? If so, rinse really well and gently pat dry. What species are you trying?
The QiaGen RNeasy kit includes a "shredder" column that is used to help disrupt plant tissues. But you do need to mechanically grind the samples very thoroughly first. Either a bead-beater or grinding in liquid nitrogen.
You want the tissue to look like a very fine powder (like flour or makeup powder).
Amy Klocko I am using switchgrass roots. We did wash and dry them before grinding and freezing them. These roots are tough to grind but we did get some with fine powder, but even those, do not give me quality RNA
Katie A S Burnette we have ground the roots as best possible, but I have not tried that kit yet. Maybe I can give it a try.
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