<html xmlns:v="urn:schemas-microsoft-com:vml" xmlns:o="urn:schemas-microsoft-com:office:office" xmlns:w="urn:schemas-microsoft-com:office:word" xmlns:m="http://schemas.microsoft.com/office/2004/12/omml" xmlns="http://www.w3.org/TR/REC-html40">
<head>
<meta http-equiv="Content-Type" content="text/html; charset=utf-8">
<meta name="Generator" content="Microsoft Word 15 (filtered medium)">
<style><!--
/* Font Definitions */
@font-face
{font-family:"Cambria Math";
panose-1:2 4 5 3 5 4 6 3 2 4;}
@font-face
{font-family:Calibri;
panose-1:2 15 5 2 2 2 4 3 2 4;}
/* Style Definitions */
p.MsoNormal, li.MsoNormal, div.MsoNormal
{margin:0cm;
margin-bottom:.0001pt;
font-size:12.0pt;
font-family:"Times New Roman",serif;}
a:link, span.MsoHyperlink
{mso-style-priority:99;
color:blue;
text-decoration:underline;}
a:visited, span.MsoHyperlinkFollowed
{mso-style-priority:99;
color:purple;
text-decoration:underline;}
span.hoenzb
{mso-style-name:hoenzb;}
span.EmailStyle19
{mso-style-type:personal-reply;
font-family:"Calibri",sans-serif;
color:#1F497D;}
.MsoChpDefault
{mso-style-type:export-only;
font-family:"Calibri",sans-serif;
mso-fareast-language:EN-US;}
@page WordSection1
{size:612.0pt 792.0pt;
margin:72.0pt 72.0pt 72.0pt 72.0pt;}
div.WordSection1
{page:WordSection1;}
--></style><!--[if gte mso 9]><xml>
<o:shapedefaults v:ext="edit" spidmax="1026" />
</xml><![endif]--><!--[if gte mso 9]><xml>
<o:shapelayout v:ext="edit">
<o:idmap v:ext="edit" data="1" />
</o:shapelayout></xml><![endif]-->
</head>
<body lang="EN-GB" link="blue" vlink="purple">
<div class="WordSection1">
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Modern techniques such as Next Generation Sequencing could well yield something but it will be a greater challenge from samples treated
with Mercuric based fixatives than formaldehyde as it forms stronger chemical crosslinks than formaldehyde, particularly with sulphur groups in proteins and nucleic acids. Pre-treatments to attempt to reverse the fixation reactions and remove mercury will
be key to any possible PCR success, but this will be harder the older the samples. E.g. see Hopwood (2002) in Bancroft and Gamble’s Theory and Practice of histological techniques. Julie Eklund also provides evidence of Mercury (II) chloride damage to short
amplified DNA sections in her 2007 thesis ‘The effects and preparation of conservation treatments on DNA’.
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Thus if the samples have been collected and subsequently stored in Mercury based fixative then the chance of getting usable DNA will
probably be extremely low, but if the samples were moved to a more suitable preservative after collecting then that will probably improve the chances of amplification success. I guess the only way to know is to give it a go and let us know if it works or not
– I’d certainly be interested to know either way!<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Best wishes<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US">Jules<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Prif Gadwraethydd, Gwyddorau Naturiol
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">Principal Conservator Natural Sciences<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><a href="mailto:julian.carter@amgueddfacymru.ac.uk"><span style="color:#0563C1">julian.carter@amgueddfacymru.ac.uk</span></a><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><a href="mailto:julian.carter@museumwales.ac.uk"><span style="color:#0563C1">julian.carter@museumwales.ac.uk</span></a><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">029 20573230<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="CY" style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D">07870448074</span><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt;font-family:"Calibri",sans-serif;color:#1F497D;mso-fareast-language:EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><b><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif">From:</span></b><span lang="EN-US" style="font-size:11.0pt;font-family:"Calibri",sans-serif"> nhcoll-l-bounces@mailman.yale.edu [mailto:nhcoll-l-bounces@mailman.yale.edu]
<b>On Behalf Of </b>Paolo Viscardi<br>
<b>Sent:</b> 08 February 2017 07:53<br>
<b>To:</b> Rachael Peart <rachaelapeart@gmail.com><br>
<b>Cc:</b> Joachim Haendel <joachim.haendel@zoologie.uni-halle.de>; nhcoll-l@mailman.yale.edu; Dirk Neumann <dirk.neumann@zsm.mwn.de>; Kareen Schnabel <Kareen.Schnabel@niwa.co.nz><br>
<b>Subject:</b> Re: [Nhcoll-l] FW: [CRUST-L:9947] Mercuric chloride and sequencing<o:p></o:p></span></p>
<p class="MsoNormal"><o:p> </o:p></p>
<div>
<p class="MsoNormal">Hi Rachael,<o:p></o:p></p>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
<div>
<p class="MsoNormal">these are both quite old references and contemporary techniques can yield usable data from much more fragmented DNA than was possible back in 2003. I expect the High Throughput Sequencing techniques being used for formalin treated material
my be applicable to specimens preserved in mercuric chloride: <a href="https://urldefense.proofpoint.com/v2/url?u=http-3A__journals.plos.org_plosone_article-3Fid-3D10.1371_journal.pone.0141579&d=CwMFaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=7Hv0d9U59ARm-uJ3xwzPGUpdSBFvTyKM0ZHs1Gr8kUQ&s=wfyTfwZHLSNzoV9KY6fhmWfvpjFkXDgHSIRmQKVcMmA&e=">
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0141579</a> <o:p></o:p></p>
</div>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
<div>
<p class="MsoNormal">Cheers,<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
<div>
<p class="MsoNormal">Paolo<o:p></o:p></p>
</div>
</div>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
<div>
<p class="MsoNormal">On 24 January 2017 at 20:14, Rachael Peart <<a href="mailto:rachaelapeart@gmail.com" target="_blank">rachaelapeart@gmail.com</a>> wrote:<o:p></o:p></p>
<blockquote style="border:none;border-left:solid #CCCCCC 1.0pt;padding:0cm 0cm 0cm 6.0pt;margin-left:4.8pt;margin-right:0cm">
<div>
<div>
<p class="MsoNormal">Thanks for the discussion,<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal">It looks like due to it's toxic nature, there is no extraction possible from animals preserved with mercuric chloride based products.<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal">Found these articles.<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal">O'Leary et al, 1994 - "The importance of fixation procedures on DNA template and its suitability for solution-phase polymerase chain reaction and PCR
<em>in situ </em>hybridisation."<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal">The Histochemical Journal 26 (4): 337 - 346.<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal">C. Schander and K.M. Halanych, 2003. "DNA, PCR and formalinized animal tissue - a short review and protocols." Organisms Diversity and Evolution 3 (3): 195 - 205.<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
<div>
<p class="MsoNormal">Thanks anyway<o:p></o:p></p>
</div>
<div>
<p class="MsoNormal"><span style="color:#888888"><o:p> </o:p></span></p>
</div>
<div>
<p class="MsoNormal"><span style="color:#888888">Rachael<o:p></o:p></span></p>
</div>
</div>
<div>
<div>
<div>
<p class="MsoNormal"><o:p> </o:p></p>
<div>
<p class="MsoNormal">On Tue, Jan 24, 2017 at 9:50 PM, Joachim Haendel <<a href="mailto:joachim.haendel@zns.uni-halle.de" target="_blank">joachim.haendel@zns.uni-halle.de</a>> wrote:<o:p></o:p></p>
<blockquote style="border:none;border-left:solid #CCCCCC 1.0pt;padding:0cm 0cm 0cm 6.0pt;margin-left:4.8pt;margin-right:0cm">
<p class="MsoNormal">Hello Kareen & Rachael,<br>
and hello Dirk,<br>
<br>
well - this is really interesting. But I can't say anything about this. To my knowledge there are no researches<br>
on this topic. Mercury(II) chloride is an effective antiseptic and disinfectant and it is corrosive. It is thinkable<br>
that it will damage the DNA. Maybe someone could try it out?! I think many museo-biologists want to know<br>
the results.<br>
<br>
Greetings<br>
Joachim<br>
<br>
<br>
<br>
---- On 24 Jan 2017 at 8:36, Dirk Neumann wrote:<br>
<br>
> Good morning Kareen & Rachael,<br>
><br>
> I am not sure if there is first hand experience how mercuric chloride<br>
> behaves exactly, but it was used as fixative especially for marine<br>
> invertebrates in the early 20th century (sublimate; cf. Piechocki &<br>
> Händel, 2007. Makroskopische Präparationstechnik, Wirbellose, 5th<br>
> Edition) and as early antibiotic (e.g. against syphilis) because of its<br>
> toxic properties. So I would assume that it does interact with tissues<br>
> and DNAs and consequently do influence sequencing results.<br>
><br>
> Perhaps Joachim can add more details ?<br>
><br>
> Hope this helps<br>
> Dirk<br>
><br>
> Am 23.01.2017 um 23:54 schrieb Kareen Schnabel:<br>
> ><br>
> > Good morning all, I thought this listserv may have some valuable input<br>
> > into my colleague´s query to the Crustacean listserv. Please see query<br>
> > below and please reply to Rachael Peart <a href="mailto:rachaelapeart@gmail.com" target="_blank">
rachaelapeart@gmail.com</a><br>
> > <mailto:<a href="mailto:rachaelapeart@gmail.com" target="_blank">rachaelapeart@gmail.com</a>>.**<br>
> ><br>
> > Thanks<br>
> ><br>
> > Kareen<br>
> ><br>
> ><br>
> > DrKareen Schnabel<br>
> > Marine Biologist<br>
> ><br>
> > *From:*<a href="mailto:crust-l-request@vims.edu" target="_blank">crust-l-request@vims.edu</a> [mailto:<a href="mailto:crust-l-request@vims.edu" target="_blank">crust-l-request@vims.edu</a>] *On<br>
> > Behalf Of *Rachael Peart<br>
> > *Sent:* Tuesday, 24 January 2017 11:49 a.m.<br>
> > *To:* <a href="mailto:crust-l@vims.edu" target="_blank">crust-l@vims.edu</a><br>
> > *Subject:* [CRUST-L:9947] Mercuric chloride and sequencing<o:p></o:p></p>
<div>
<div>
<p class="MsoNormal" style="margin-bottom:12.0pt">> ><br>
> > Hi all,<br>
> ><br>
> > I have come across some amphipod samples that were captured in traps<br>
> > using mercuric chloride.<br>
> ><br>
> > I am curious to know if this chemical can affect molecular sequencing<br>
> > - ie COI or 18S?<br>
> ><br>
> > Does it completely destroy the amounts, or do you get traces, or no<br>
> > damage?<br>
> ><br>
> > Thanks in advance<br>
> ><br>
> > Rachael Peart<br>
> ><br>
> ><br>
<br>
<br>
<o:p></o:p></p>
</div>
</div>
<p class="MsoNormal" style="margin-bottom:12.0pt">--<br>
Joachim Haendel<br>
Center of Natural Sciences Collections<br>
of the Martin-Luther-University<br>
- Entomological Collection -<br>
<br>
Domplatz 4<br>
D-06099 Halle (Saale)<br>
Germany<br>
<br>
Phone: <a href="tel:%2B49%20345%20-%2055%2026%20447" target="_blank">+49 345 - 55 26 447</a><br>
Fax: <a href="tel:%2B49%20345%20-%2055%2027%20248" target="_blank">+49 345 - 55 27 248</a><br>
Email: <a href="mailto:joachim.haendel@zns.uni-halle.de" target="_blank">joachim.haendel@zns.uni-halle.de</a><br>
<br>
<o:p></o:p></p>
</blockquote>
</div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
</div>
</div>
<p class="MsoNormal" style="margin-bottom:12.0pt"><br>
_______________________________________________<br>
Nhcoll-l mailing list<br>
<a href="mailto:Nhcoll-l@mailman.yale.edu">Nhcoll-l@mailman.yale.edu</a><br>
<a href="http://mailman.yale.edu/mailman/listinfo/nhcoll-l" target="_blank">http://mailman.yale.edu/mailman/listinfo/nhcoll-l</a><br>
<br>
_______________________________________________<br>
NHCOLL-L is brought to you by the Society for the Preservation of<br>
Natural History Collections (SPNHC), an international society whose<br>
mission is to improve the preservation, conservation and management of<br>
natural history collections to ensure their continuing value to<br>
society. See <a href="https://urldefense.proofpoint.com/v2/url?u=http-3A__www.spnhc.org&d=CwMFaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=7Hv0d9U59ARm-uJ3xwzPGUpdSBFvTyKM0ZHs1Gr8kUQ&s=8QklzJEgCsu-wWTqUkDgjGCUYSt_pyI3TZ8Ua2aAxb4&e=" target="_blank">
http://www.spnhc.org</a> for membership information.<br>
Advertising on NH-COLL-L is inappropriate.<o:p></o:p></p>
</blockquote>
</div>
<p class="MsoNormal"><o:p> </o:p></p>
</div>
</div>
<p> </p>
<p> </p>
<p><span lang="CY" style="font-family: 'Arial',sans-serif;">YMWADIAD<br /> Rydym yn croesawu gohebiaeth yn Gymraeg a Saesneg, ac yn sicrhau y byddwn yn cyfathrebu â chi yn eich iaith ddewisol, boed yn Gymraeg, Saesneg neu’r ddwy, dim ond i chi ein hysbysu. Ni fydd gohebu yn Gymraeg yn peri oedi.</span></p>
<p> </p>
<p><a name="cysill"></a><span lang="CY" style="font-family: 'Arial',sans-serif;">Mae pob neges e-bost a anfonir at neu gan Amgueddfa Cymru yn cael ei <br /> sganio gan systemau diogelwch awtomatig. Sganiwyd y neges hon am firysau cyn ei hanfon, ond dylech hefyd wirio bod y neges, a phob atodiad ynddi, yn rhydd o firysau cyn ei defnyddio. Nid ydym yn derbyn cyfrifoldeb am unrhyw golled neu ddifrod o ganlyniad i agor y neges neu unrhyw atodiadau. Gall y neges hon ac unrhyw ffeiliau a atodir ynddi gynnwys gwybodaeth gyfrinachol a fwriadwyd ar gyfer y derbynnydd yn unig. Os ydych chi wedi derbyn y neges trwy gamgymeriad, hysbyswch ni a dileu’r neges. Safbwyntiau personol yr awdur a fynegir yn y neges hon, ac nid ydynt o reidrwydd yn cynrychioli safbwyntiau Amgueddfa Cymru. Nid ydym yn derbyn cyfrifoldeb am unrhyw wallau, llygredd neu esgeulustod a allai godi wrth drosglwyddo'r neges hon.</span></p>
<p> </p>
<p><span lang="CY" style="font-family: 'Arial',sans-serif;"> </span></p>
<p> </p>
<p style="margin-right: -2.85pt;"><span lang="CY" style="font-family: 'Arial',sans-serif;">DISCLAIMER</span></p>
<p> </p>
<p><span lang="CY" style="font-family: 'Arial',sans-serif;">We welcome correspondence in Welsh and English, and we will ensure that we communicate with you in the language of your choice, whether that’s English, Welsh or both if you let us know which you prefer. Corresponding in Welsh will not lead to any delay.</span></p>
<p> </p>
<p><span style="font-family: 'Arial',sans-serif; mso-ansi-language: EN-GB;">E-mail to and from Amgueddfa Cymru is scanned by automated security systems. This message was scanned for viruses before transmission, but you should also satisfy yourself that the message, and all attachments, are virus-free before use. We can accept no responsibility for any loss or damage that might arise from opening the message or any attachments. This message and any files transmitted with it may contain confidential information intended only for the recipient. If you receive the message by mistake please inform us and delete it. The views expressed in this message are the personal views of the author and may not necessarily represent those of Amgueddfa Cymru. We accept no liability for any errors, corruption or omissions that might arise in transmission of this message.</span></p>
<p> </p>
<p> </p>
<hr />
<p> </p>
<p><br />Scanned by FuseMail.</p></body>
</html>