Removal of mitochondria RNA from total RNA? - (Mar/08/2008 )

Hi everyone,

I am working on some whole genome sequencing and am finding that a lot of sequencing is coming from mitochondria.

Is there a way to remove mitochondrial RNA from total RNA? Perhaps a bead-based system?

I take total RNA and RiboMinus it to remove ribosomal RNA, but I need to see about getting rid of mito RNA too.

Thank!

Maybe it will be easy to remove mitochondia before starting the RNA extraction. In plants there are some protocols to isolate mitochondria, so maybe you could use one of those to get yourself rid of them.

P.S. I'm not sure whether this works all that well, I've never tried it. But I don't know any method to isolate mitochondrial RNA from the rest, so...

You should be able to make custom "capture beads" which are solid phase agarose or other beads with oligos that are complementary to the sequences you want to eliminate. These should be easy to make by ordering biotinylated complementary oligos and linking them to streptavidin beads. Then, you capture the undesired sequences by annealing with the beads and filtering them out. This is what is done with rRNA capture beads for reducing ribosomal RNA in cDNA experiments.