[Nhcoll-l] Use of denatured ethanol for short term storage of molecular samples

[Dirk Neumann]

Hi Nick,

the question is which denaturants are included, because additives behave 
differently; also the integument of target organisms influences 
dehydration of tissues.

An alternative (not tested myself) might be strong liquors (if 
available) or to mix the denatured EtOH with strong liquor to reduce 
potentially negative effects of denaturants, but you would need to 
exchange the fluid surely 2-3 times to avoid hydrolysis and guarantee 
fast dehydration of tissues in order to rapidly preserve the DNA (tissue 
: fluid volume is decisive).

Another alternative (depending on outside temperatures or availability 
of ice) would be to freeze the frogs in the field to tissue them, or to 
rapidly sun dry smaller amounts of tissues (but you would need to make 
sure that dried tissues stay absolutely dry to avoid dehydration).

If denaturants interfere with the helix you would have smaller fragments 
of course, but if you extract tissues fast after returning from the 
field the yield still may be good even with denatured ethanol. But as 
said: key is to stop hydrolysis and autolysis by inactivating enzymes or 
dehydrating tissues fast.

Hopes this gives you some inspirations

Best wishes
Dirk




Am 18.04.2019 um 23:03 schrieb Nick Cairns:
> Hello everyone,
> I'm seeking guidance on reagents. I'm trying to collect chorus frogs 
> from across western Canada (whole and toe clips). These samples will 
> be likely be extracted using phenol/chloroform then ethanol (EtOH) 
> precipitation to tidy them up. Downstream they'll be used for mtDNA 
> (Sanger) and genomic (ddRAD) protocols. The issue is, I am currently 
> in rural Saskatchewan and only have denatured EtOH (Fisherbrand 
> Histoprep 95%) available to me.  I understand that the additives in 
> some denatured EtOH can cause issues downstream but has anyone ever 
> used it for short term storage then replaced it later with anhydrous 
> to remove the additives? Can these additives be reduced from the 
> tissues after the fact?
> Any insights would be most welcome.
> Thank you,
> Nick
>
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-- 
Dirk Neumann

Tel: 089 / 8107-111
Fax: 089 / 8107-300
*new email: neumann(a)snsb.de*

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Tel: +49-89-8107-111
Fax: +49-89-8107-300
*new email: neumann(a)snsb.de*

postal address:

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