How do I extract RNA from small amounts of tissue with high quality using Tri-reagent?

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Melissa M McGovern

I have tried several times now and I've even tried taking less of the upper phase trying to avoid contamination, but it seems hit or miss. I guess it will just take practice.

Bosch Steffi

Hello Melissa,

How small is your tissue sample? be careful to dissociate the tissue thoroughly. Precipitation of small amounts of nucleic acids may be a limiting step. If suitable with your downstream applications, adding a carrier like yeast tRNA or linear polyacrylamide might help. Good luck, Steffi

I'm not sure the exact weight, but my typical RNA yield is about 200ng/ul re-suspended in 20ul. I use the beads to homogenize the sample in the tri-reagent . I currently precipitate in -20 for about 48 hours, though a colleague has suggested trying to precipitate in -80. I'm using the RNA for real time qPCR.