RNA from MMA embedded calcified bone - (Oct/29/2009 )
Does anyone have experience with isolating RNA from MMA embedded calcified bone?
We are working a lot with MMA embedded bone and metal implants. We got the idea that it could be very nice to be able to isolate RNA from specific sites around the implant with laser microdissection. We are embedding the bone in MMA (Methylmethacrylate) and I can't seem to find any publications about this. So I don't know if it is possible to get RNA of a resonable quality out from MMA embedded material.
We've tried this about 5 years ago, including laser-microdissection, but the RNA was of pretty lousy quality. The best RIN we got from the bioanalyzer was 4.1/10, and most of it was on the 2-3 range and not useable. We tried pre-extracted the MMA with acetone before microdissetion, but did not make much difference, and more often dissolved the PET film on the LMD slide. What we did not do though was a Proteinase K digestion, now commonplace in DNA/RNA recovery from FFPE tissue (I believe Qiagen makes a kit), so I can't help but assume this might improve things in an MMA prep. Would be interesting to know if that would help, but we've since moved on to greener pastures. Good luck, and let us know how it goes.