[Nhcoll-l] FW: [CRUST-L:9947] Mercuric chloride and sequencing

Wed Feb 8 10:50:36 EST 2017

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 Gambleâ€™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 â€˜The effects and preparation of conservation treatments on DNAâ€™.

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 â€“ Iâ€™d certainly be interested to know either way!

Best wishes

Jules

Prif Gadwraethydd, Gwyddorau Naturiol
Principal Conservator Natural Sciences
julian.carter at amgueddfacymru.ac.uk<mailto:julian.carter at amgueddfacymru.ac.uk>
julian.carter at museumwales.ac.uk<mailto:julian.carter at museumwales.ac.uk>
029 20573230
07870448074

From: nhcoll-l-bounces at mailman.yale.edu [mailto:nhcoll-l-bounces at mailman.yale.edu] On Behalf Of Paolo Viscardi
Sent: 08 February 2017 07:53
To: Rachael Peart <rachaelapeart at gmail.com>
Cc: Joachim Haendel <joachim.haendel at zoologie.uni-halle.de>; nhcoll-l at mailman.yale.edu; Dirk Neumann <dirk.neumann at zsm.mwn.de>; Kareen Schnabel <Kareen.Schnabel at niwa.co.nz>
Subject: Re: [Nhcoll-l] FW: [CRUST-L:9947] Mercuric chloride and sequencing

Hi Rachael,

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: https://urldefense.proofpoint.com/v2/url?u=http-3A__journals.plos.org_plosone_article-3Fid-3D10.1371_journal.pone.0141579&d=CwIGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=LpjxosJg1wwlI83aSVVxHtSncR8Qg1P84yJfVz6O0tk&s=f6Vl5K0e3nX3S_74alp5oVEiGTcRh9YRP04PT9e_IE4&e= <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=>

Cheers,

Paolo

On 24 January 2017 at 20:14, Rachael Peart <rachaelapeart at gmail.com<mailto:rachaelapeart at gmail.com>> wrote:
Thanks for the discussion,
It looks like due to it's toxic nature, there is no extraction possible from animals preserved with mercuric chloride based products.
Found these articles.
O'Leary et al, 1994 - "The importance of fixation procedures on DNA template and its suitability for solution-phase polymerase chain reaction and PCR in situ hybridisation."
The Histochemical Journal 26 (4): 337 - 346.
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.

Thanks anyway

Rachael

On Tue, Jan 24, 2017 at 9:50 PM, Joachim Haendel <joachim.haendel at zns.uni-halle.de<mailto:joachim.haendel at zns.uni-halle.de>> wrote:
Hello Kareen & Rachael,
and hello Dirk,

well - this is really interesting. But I can't say anything about this. To my knowledge there are no researches
on this topic. Mercury(II) chloride is an effective antiseptic and disinfectant and it is corrosive. It is thinkable
that it will damage the DNA. Maybe someone could try it out?! I think many museo-biologists want to know
the results.

Greetings
Joachim

---- On 24 Jan 2017 at 8:36, Dirk Neumann wrote:

> Good morning Kareen & Rachael,
>
> I am not sure if there is first hand experience how mercuric chloride
> behaves exactly, but it was used as fixative especially for marine
> invertebrates in the early 20th century (sublimate; cf. Piechocki &
> HÃ¤ndel, 2007. Makroskopische PrÃ¤parationstechnik, Wirbellose, 5th
> Edition) and as early antibiotic (e.g. against syphilis) because of its
> toxic properties. So I would assume that it does interact with tissues
> and DNAs and consequently do influence sequencing results.
>
> Perhaps Joachim can add more details ?
>
> Hope this helps
> Dirk
>
> Am 23.01.2017 um 23:54 schrieb Kareen Schnabel:
> >
> > Good morning all, I thought this listserv may have some valuable input
> > into my colleagueÂ´s query to the Crustacean listserv. Please see query
> > below and please reply to Rachael Peart rachaelapeart at gmail.com<mailto:rachaelapeart at gmail.com>
> > <mailto:rachaelapeart at gmail.com<mailto:rachaelapeart at gmail.com>>.**
> >
> > Thanks
> >
> > Kareen
> >
> >
> > DrKareen Schnabel
> > Marine Biologist
> >
> > *From:*crust-l-request at vims.edu<mailto:crust-l-request at vims.edu> [mailto:crust-l-request at vims.edu<mailto:crust-l-request at vims.edu>] *On
> > Behalf Of *Rachael Peart
> > *Sent:* Tuesday, 24 January 2017 11:49 a.m.
> > *To:* crust-l at vims.edu<mailto:crust-l at vims.edu>
> > *Subject:* [CRUST-L:9947] Mercuric chloride and sequencing
> >
> > Hi all,
> >
> > I have come across some amphipod samples that were captured in traps
> > using mercuric chloride.
> >
> > I am curious to know if this chemical can affect molecular sequencing
> > - ie COI or 18S?
> >
> > Does it completely destroy the amounts, or do you get traces, or no
> > damage?
> >
> > Thanks in advance
> >
> > Rachael Peart
> >
> >

--
Joachim Haendel
Center of Natural Sciences Collections
of the Martin-Luther-University
- Entomological Collection  -

Domplatz 4
D-06099 Halle (Saale)
Germany

Phone:  +49 345 - 55 26 447<tel:%2B49%20345%20-%2055%2026%20447>
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