Protecting RNA in the jungle

Deanne Bell dbell at fresno.ars.usda.gov
Tue Aug 23 11:03:24 EST 2005


Hi all

Sorry for the late response, but I heartily agree that you should not
crush the material until you return.  Just removing your flower from the
plant should start a series of cell death responses that could damage
nucleic acids. Crushing the fresh tissue will release a flood of acids
and waste products that will decrade the nucleic acids, let alone all
the RNases that will degrade your RNA within hours at ambient
temperatures.

I would try RNAlater (from Ambion, but no affiliation) for 1 day at
ambient temp and get the samples into frig or freezer within 1 day.  I
don't know if you can get RNA from dried tissue, but if your trip into
the jungle will be for a prolonged period of time without any
refrigeration, you could probably dessicate the tissue samples and get
DNA at a later time.  I have also isolated good quality DNA from plants
and insects stored in ETOH.  I think you may need to consider working
with DNA.

Hope this helps
Deanne Bell
USDA Agricultural Research Service
Crop Diseases, Pests & Genetics
9611 South Riverbend Avenue
Parlier, CA  93648
voice  (559) 596-2806
fax      (559) 596-2897
dbell at fresno.ars.usda.gov
 
 
 

>-----Original Message-----
>From: methods-bounces at oat.bio.indiana.edu 
>[mailto:methods-bounces at oat.bio.indiana.edu] On Behalf Of Bo Johansen
>Sent: Tuesday, August 16, 2005 1:00 AM
>To: methods at magpie.bio.indiana.edu
>Subject: Protecting RNA in the jungle
>
>Hi
>
>I am going on a trip to collect RNA samples from flower buds 
>in the jungle of plants that cannot be cultivated. It is 
>impossible to bring liquid nitrogen on the trip, and I won't 
>get into a lab for 2-3 weeks. 
>What do I do.
>
>I have thought of collecting the material in Eppendorph tubes, 
>add 1 ml og 2.5 M LiCl and crush the material with a pistil. 
>Close the tube and leave it until I get to a lab
>
>Or
>
>Collecting the material in an Eppendorph tube, add 1 ml og  4M 
>guanidine isothiocyanate, 25mM sodium citrate, pH7, 0.5% 
>sarcosyl, 100mM b-mercaptoethanolm, crush the material and 
>leave it until I get to a lab.
>
>Will any of these methods leave the RNA intact or are there 
>other methods?
>
>Bo
>
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