[Coral-List] RNA sampling in the field

sarah davies daviessw at gmail.com
Thu Nov 10 17:43:00 EST 2016


Hello listers,

I'm writing to let you all know about the recent success I have had
sampling RNA in the field. I was doing fieldwork at the Flower Garden Banks
and I was keen to get RNA samples, however the depths of FGB are limiting
and you must be able to preserve while under water. So I tried this new
method and I am happy to report that it works beautifully as long as you
have a trusty buddy!

Here are the steps I followed:
1. Prefill 15ml conical tubes with 200 proof EtOH making sure there were no
air bubbles.
2. Sampling requires two dedicated divers: one to sample from the colony
and the other to carefully open the 15ml tube upside down.
3. The sample is quickly transferred onto the center of the conical lid
(needs to be a small tissue sample so you can easily close). Then the tube
is quickly sealed (some EtOH leaks out as soon as you open so it is not
perfect)
4. Shake vigorously.
5. Once at the surface, transfer sample to a new 2ml cryo tube prefilled
with fresh 200 proof EtOH.
6. Keep as cold as possible until RNA isolation. I kept in a standard
freezer, transported on blue ice and then stored at -80 until isolation.

Here is the photo of the isolations on a gel. Nice RNA bands and really
good yields!
https://www.instagram.com/swdavies03/?hl=en

I hope this protocol will be helpful for those of you interested in
sampling RNA in the field.

Cheers
Sarah

-- 
Sarah W. Davies M.Sc. Ph.D.
Simons Foundation Fellow of the Life Sciences Research Foundation
Postdoctoral Research Associate
Department of Marine Sciences
University of North Carolina at Chapel Hill
https://sarahwdavies.wordpress.com/
daviessw at gmail.com
1-512-609-9134


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