Isolating RNA from frozen tissues kept in vials - (Jun/13/2008 )
Somebody sent me frozen tissues kept in 1.5 ml screw-cap vials and I need to isolate RNA from those tissues.
My problem is, I`m not sure how I can take the tissues out from the tube without thawing it.
I`m thinking to hit the tube with hammer, then dip the tissues immediately to liquid nitrogen, and place it in a ziplock plastic bag.
Then I will put it back at -80 C, and crash it with a hammer to make it into smaller pieces before I start the isolation.
Oh btw, I will do the RNA isolation on different day with the "frozen tissue preparation".
But I`m really not sure about this....
Do you have any better idea?
Please..I need your suggestion
Thanks!
I used to collect patients biopsies and preserve in a liquid called RNAi prior to RNA extraction. check if u could thaw the samples in it.
with a mortar and pestle that resist LN2. Cool the mortar and pestle leaving at least 24h in freezer (the change in temp will crack it). Set all before taking out the samples and the mortar. Add the sample to the mortar and then add LN2 and hit with the pestle until you have a fine powder.
My problem is, I`m not sure how I can take the tissues out from the tube without thawing it.
I`m thinking to hit the tube with hammer, then dip the tissues immediately to liquid nitrogen, and place it in a ziplock plastic bag.
Then I will put it back at -80 C, and crash it with a hammer to make it into smaller pieces before I start the isolation.
Oh btw, I will do the RNA isolation on different day with the "frozen tissue preparation".
But I`m really not sure about this....
Do you have any better idea?
Please..I need your suggestion
Thanks!
1. Add trizol to it, put a sterile stick (bent at the end) in the trizol, freeze it in -70, take out the candy, after a few minutes at RT, or hand heat. Seriously.
2. Add trizol, leave the tube on ice, once it thaws, remove the tissue and trizol for homogenization.
2. I always use kontes microfuge homogenizer for homogenization. In which case, you can directly homogenize in the tube if total sample is less than 500 microl.
thanks for your ideas 
although actually I managed to isolate the RNA from those frozen tissues in vials.
I dipped the vial in liquid nitrogen, took it out, put it in a zipped lock bag, hit with hammer, then the tissues became smaller pieces.
The isolated RNA was just fine and my real time PCR was also fine.
By the way, people in my lab never use any mortar and pestle and I think it`s because we use a multi-shocker machine?
again, thank you so much guys!