<html>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=UTF-8">
</head>
<body text="#000000" bgcolor="#FFFFFF">
<div class="moz-cite-prefix">Hi Nick,</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">the question is which denaturants are
included, because additives behave differently; also the
integument of target organisms influences dehydration of tissues.</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">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).</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">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).</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">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.</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">Hopes this gives you some inspirations</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">Best wishes</div>
<div class="moz-cite-prefix">Dirk<br>
</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix"><br>
</div>
<div class="moz-cite-prefix">Am 18.04.2019 um 23:03 schrieb Nick
Cairns:<br>
</div>
<blockquote type="cite"
cite="mid:CAB3OAaMruqzxexwj9RmgpAxaWsUn+yJSCYpBHN_sjyuAB1jSow@mail.gmail.com">
<meta http-equiv="content-type" content="text/html; charset=UTF-8">
<div dir="ltr">
<div dir="ltr">
<div>Hello everyone,</div>
<div>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 <span class="gmail-st">anhydrous</span> to remove the
additives? Can these additives be reduced from the tissues
after the fact? <br>
</div>
<div>Any insights would be most welcome. <br>
</div>
<div>Thank you,</div>
<div>Nick<br>
</div>
</div>
</div>
<br>
<fieldset class="mimeAttachmentHeader"></fieldset>
<pre class="moz-quote-pre" wrap="">_______________________________________________
Nhcoll-l mailing list
<a class="moz-txt-link-abbreviated" href="mailto:Nhcoll-l@mailman.yale.edu">Nhcoll-l@mailman.yale.edu</a>
<a class="moz-txt-link-freetext" href="https://mailman.yale.edu/mailman/listinfo/nhcoll-l">https://mailman.yale.edu/mailman/listinfo/nhcoll-l</a>
_______________________________________________
NHCOLL-L is brought to you by the Society for the Preservation of
Natural History Collections (SPNHC), an international society whose
mission is to improve the preservation, conservation and management of
natural history collections to ensure their continuing value to
society. See <a class="moz-txt-link-freetext" href="http://www.spnhc.org">http://www.spnhc.org</a> for membership information.
Advertising on NH-COLL-L is inappropriate.
</pre>
</blockquote>
<p><br>
</p>
<pre class="moz-signature" cols="72">--
Dirk Neumann
Tel: 089 / 8107-111
Fax: 089 / 8107-300
*new email: neumann(a)snsb.de*
Postanschrift:
Staatliche Naturwissenschaftliche Sammlungen Bayerns
Zoologische Staatssammlung München
Dirk Neumann, Sektion Ichthyologie / DNA-Storage
Münchhausenstr. 21
81247 München
Besuchen Sie unsere Sammlung:
<a class="moz-txt-link-freetext" href="http://www.zsm.mwn.de/sektion/ichthyologie-home/">http://www.zsm.mwn.de/sektion/ichthyologie-home/</a>
---------
Dirk Neumann
Tel: +49-89-8107-111
Fax: +49-89-8107-300
*new email: neumann(a)snsb.de*
postal address:
Bavarian Natural History Collections
The Bavarian State Collection of Zoology
Dirk Neumann, Section Ichthyology / DNA-Storage
Muenchhausenstr. 21
81247 Munich (Germany)
Visit our section at:
<a class="moz-txt-link-freetext" href="http://www.zsm.mwn.de/sektion/ichthyologie-home/">http://www.zsm.mwn.de/sektion/ichthyologie-home/</a>
</pre>
</body>
</html>