Right solvent to dissolve ampicillin ?

Eva Freisinger freisinger at aci.unizh.ch
Mon Mar 6 02:05:28 EST 2006


Hi, You will NEVER be able to make a 100mg/mL stock solution of pure amp in 
pure H2O! That is because pure amp is an acid and to dissolve it, you have 
to add approx. 1 equivalent of NaOH as well. What you guys are talking 
about is the sodium salt of amp, which you normally buy... And this is 
READILY soluble! Trust me, I tryed to dissolve pure amp in pure H2O once 
myself - before I remembered my chemistry courses... Eva

At 13:28 04.03.2006, Siddhartha Parida wrote:
>My ampicillin sod.salt has expired (exp date june 2004 ) will it be
>viable still ? therefore  i am trying with ampicillin capsules found
>in medicine stores , but after getting the calculation right it is not
>dissolving in water nor alcohol , can anyone suggest me the right
>solvent
>
>I will be using this stock solution of ampicillin in LBA medium for a
>alpha complementation protocol
>
>On 2/28/06, methods-request at oat.bio.indiana.edu
><methods-request at oat.bio.indiana.edu> wrote:
> > Send Methods mailing list submissions to
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> > than "Re: Contents of Methods digest..."
> >
> >
> > Today's Topics:
> >
> >   1. Re: protein heterodimer seperated during purification with
> >      superdex200 (DK)
> >   2. Primers, DDWater, TE Buffer (Alvin Alejandrino)
> >   3. Re: TUNEL (DK)
> >   4. Mass Ladder (ali)
> >   5. Re: Primers, DDWater, TE Buffer (Christian Praetorius)
> >   6. looking for non radioactive kinase assays (Carlo Zambonelli)
> >   7. the condition of electroporating MDBK (Zhou Xiaoyan)
> >   8. DNA digest in gel (cheetah.zao at gmail.com)
> >   9. Re: Primers, DDWater, TE Buffer (GysdeJongh)
> >  10. Re: Primers, DDWater, TE Buffer (Rajesh Kumar Singh)
> >  11. Re: Primers, DDWater, TE Buffer (Christian Praetorius)
> >  12. Re: Primers, DDWater, TE Buffer (Christian Praetorius)
> >
> >
> > ----------------------------------------------------------------------
> >
> > Message: 1
> > Date: Mon, 27 Feb 2006 14:21:51 GMT
> > From: dk at no.email.thankstospam.net (DK)
> > Subject: Re: protein heterodimer seperated during purification with
> >        superdex200
> > To: methods at net.bio.net
> > Message-ID: <dtv1tv$kto$1 at news.doit.wisc.edu>
> >
> > In article <1141031176.320387.225840 at t39g2000cwt.googlegroups.com>, 
> "cheetah.zao at gmail.com" <cheetah.zao at gmail.com> wrote:
> > >hi,everyone
> > >
> > >I have been working on purifying a protein heterodimer using affinity
> > >column(one of target protein has a GST tag) and superdex200.
> > >
> > >the first part went well, I got two bands in SDS gel, however, I met
> > >something weird when using superdex200 to purify the eluents of the
> > >affinity column: there were two main peaks,with one of my target
> > >protein seperately.
> > >
> > >Is it quit unusual for superdex200 to drag protein complex apart,
> > >
> > >has anyone met this too?
> >
> > Alll the time. It's not that gel-filtration matrix "drags protein complex
> > apart" but the fact that your complex is of relatively low affinity.
> > For those, hal-life time of the complex is small (say, minutes instead of
> > hours or days), thus if the the two proteins are considerably different
> > in size, they eventually separate.
> >
> > The situation is compeltely different in affinity chromatography,
> > where one of the components is always at high concentration
> > (> or >>Kd, ~ 0.1 mM is typical).
> >
> > DK
> >
> >
> > ------------------------------
> >
> > Message: 2
> > Date: Mon, 27 Feb 2006 10:18:44 -0800
> > From: "Alvin Alejandrino" <aalejandrino at nhm.org>
> > Subject: Primers, DDWater, TE Buffer
> > To: <methods at magpie.bio.indiana.edu>
> > Message-ID: <EE6DFADE289313489E832D79B2C5E45D0242CD19 at hermes.nhm.lac>
> > Content-Type: text/plain;       charset="iso-8859-1"
> >
> > Hey everyone,
> >
> > Normally, I use TE Buffer to dilute and resuspend primers from the 
> manufacturer.  Some people have told me that Double Distilled Water is an 
> okay and easy way to dilute the primers.  Any thoughts?  It seems way too 
> easy.  What problems will I encounter?
> >
> >
> > ------------------------------
> >
> > Message: 3
> > Date: Mon, 27 Feb 2006 14:29:09 GMT
> > From: dk at no.email.thankstospam.net (DK)
> > Subject: Re: TUNEL
> > To: methods at net.bio.net
> > Message-ID: <dtv2bm$kto$2 at news.doit.wisc.edu>
> >
> > In article <EpBMf.17322$yK1.5244 at news-server.bigpond.net.au>, "newsnet 
> customer" <customer at newsnet.com> wrote:
> > >This is a multi-part message in MIME format.
> > >
> > >------=_NextPart_000_0011_01C63BE9.1744EE50
> > >Content-Type: text/plain;
> > >        charset="iso-8859-1"
> > >Content-Transfer-Encoding: quoted-printable
> >
> > Please only post plaintext messages on Usenet. Thank you.
> >
> > >Is anyone familiar with TUNEL assay.
> > >All i know is that it detects fragmented DNA.
> > >Anyone know how it works?
> >
> > It works by incorporating labeled (usually fluorescent)
> > nucleotide (usuallly dUTP) to the termini of DNA by
> > a specific enzyme (TdT terminal transferase).
> > The more free ends (as in apoprosis where DNA is
> > fragmented), the higher incorporation, the higher
> > signal.
> >
> > DK
> >
> >
> > ------------------------------
> >
> > Message: 4
> > Date: 27 Feb 2006 13:09:03 -0800
> > From: "ali" <atahery at gmail.com>
> > Subject: Mass Ladder
> > To: methods at net.bio.net
> > Message-ID: <1141074543.829968.265400 at p10g2000cwp.googlegroups.com>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> > Do we have to buy the mass ladder all the time or it is possible to
> > make our own. Except spect and Gel,  is there any reliable method is
> > quantifying DNA?
> >
> >
> >
> > ------------------------------
> >
> > Message: 5
> > Date: Mon, 27 Feb 2006 21:37:16 +0000
> > From: Christian Praetorius <prae at gmx.net>
> > Subject: Re: Primers, DDWater, TE Buffer
> > To: methods at net.bio.net
> > Message-ID: <46h9obFbaabjU1 at individual.net>
> > Content-Type: text/plain; charset=us-ascii
> >
> > "Alvin Alejandrino" <aalejandrino at nhm.org> wrote:
> >
> > >Normally, I use TE Buffer to dilute and resuspend primers from the 
> manufacturer.  Some people have told me that Double Distilled Water is an 
> okay and easy way to dilute the primers.  Any thoughts?  It seems way too 
> easy.  What problems will I encounter?
> >
> > I think, it doesn't matter. TE may be better because it is buffered
> > and my complex ions, which are necessary for nucleases. I always
> > dissolve and dilute my primers in double distilled water and this
> > worked always.
> >
> > Christian
> >
> > --
> >        [X] <-- nail here for new monitor
> >
> >
> > ------------------------------
> >
> > Message: 6
> > Date: Mon, 27 Feb 2006 19:06:53 -0500
> > From: "Carlo Zambonelli" <zambonel at gmail.com>
> > Subject: looking for non radioactive kinase assays
> > To: methods at magpie.bio.indiana.edu
> > Message-ID:
> >        <ad3c93330602271606v59d9f338kca6bf1071bb0c1a1 at mail.gmail.com>
> > Content-Type: text/plain; charset=ISO-8859-1
> >
> > Dear all,
> > I am looking for a non radioactive universal kinase assay kit. I found many
> > companies selling kits apparently suitable for this application: I would
> > like to receive recommendations/suggestions from first hand users of any
> > such kit. From your experience, what is the sensitivity compared to
> > radioactive methods?
> > Thanks,
> > Carlo
> >
> >
> > ------------------------------
> >
> > Message: 7
> > Date: Tue, 28 Feb 2006 10:37:07 +0800
> > From: "Zhou Xiaoyan" <zxy780405 at gmail.com>
> > Subject: the condition of electroporating MDBK
> > To: methods at magpie.bio.indiana.edu
> > Message-ID: <30463dd60602271837j4e26f01k at mail.gmail.com>
> > Content-Type: text/plain; charset=ISO-8859-1
> >
> > Hi
> >    I'm from China, I have found the electroporation of MDBK on internet.
> > Can you tell me the condition of electroporation? Thank you!
> >
> > your oversea friend
> >
> > Xiaoyan Zhou
> >
> >
> > ------------------------------
> >
> > Message: 8
> > Date: 28 Feb 2006 00:46:52 -0800
> > From: "cheetah.zao at gmail.com" <cheetah.zao at gmail.com>
> > Subject: DNA digest in gel
> > To: methods at net.bio.net
> > Message-ID: <1141116412.101620.147830 at u72g2000cwu.googlegroups.com>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> > hi  everyone, I'm new to Molecular biology.
> > In order to reduce unnecessary yield loss, I would like to  digest DNA
> > in normal Agarose gel 1%, should I make some adjustment of the reagents
> > compared to auqatic digestion, or anyone who is familiar with it has
> > other advices?
> >
> >
> > zhao
> >
> >
> >
> > ------------------------------
> >
> > Message: 9
> > Date: Tue, 28 Feb 2006 16:06:08 +0100
> > From: "GysdeJongh" <jongh711 at planet.nl>
> > Subject: Re: Primers, DDWater, TE Buffer
> > To: methods at net.bio.net
> > Message-ID: <440466dd$0$2015$ba620dc5 at text.nova.planet.nl>
> >
> > "Christian Praetorius" <prae at gmx.net> wrote in message
> > news:46j1piFbdg6eU1 at individual.net...
> > > Christian Praetorius <prae at gmx.net> wrote:
> > >
> > >>I always
> > >>dissolve and dilute my primers in double distilled water and this
> > >>worked always.
> > >
> > > One more thing: I always dilute the primer stocks to 100pmol and make
> > > a 10pmol dilution for use in PCR.
> >
> > Hi,
> > I  _think_  you mean 100 pmol / microliter , which is 100 micromolar or 100
> > micromol / liter
> > To avoid the greek alphabet  :)
> > hth
> > Gys
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 10
> > Date: Tue, 28 Feb 2006 10:05:12 +0100
> > From: Rajesh Kumar Singh <rsingh at EMBL-Hamburg.DE>
> > Subject: Re: Primers, DDWater, TE Buffer
> > To: methods at magpie.bio.indiana.edu
> > Message-ID:
> > 
> <Pine.SGI.4.44.0602281002300.1986127-100000 at butthead.embl-hamburg.de>
> > Content-Type: TEXT/PLAIN; charset=US-ASCII
> >
> > usually its not a problem in distilled water. but its good to
> > keep track of pH of water being used.
> >
> > Rajesh Kumar Singh
> > European Molecular Biology Laboratory
> > C/o DESY, Notkestrasse 85, Geb. 25A
> > 22603 Hamburg, Germany
> > E-mail: rsingh at embl-hamburg.de
> >
> > On Mon, 27 Feb 2006, Christian Praetorius wrote:
> >
> > > "Alvin Alejandrino" <aalejandrino at nhm.org> wrote:
> > >
> > > >Normally, I use TE Buffer to dilute and resuspend primers from the 
> manufacturer.  Some people have told me that Double Distilled Water is an 
> okay and easy way to dilute the primers.  Any thoughts?  It seems way too 
> easy.  What problems will I encounter?
> > >
> > > I think, it doesn't matter. TE may be better because it is buffered
> > > and my complex ions, which are necessary for nucleases. I always
> > > dissolve and dilute my primers in double distilled water and this
> > > worked always.
> > >
> > > Christian
> > >
> > > --
> > >         [X] <-- nail here for new monitor
> > > _______________________________________________
> > > Methods mailing list
> > > Methods at net.bio.net
> > > http://www.bio.net/biomail/listinfo/methods
> > >
> >
> >
> >
> > ------------------------------
> >
> > Message: 11
> > Date: Tue, 28 Feb 2006 13:33:42 +0000
> > From: Christian Praetorius <prae at gmx.net>
> > Subject: Re: Primers, DDWater, TE Buffer
> > To: methods at net.bio.net
> > Message-ID: <46j1piFbdg6eU1 at individual.net>
> > Content-Type: text/plain; charset=us-ascii
> >
> > Christian Praetorius <prae at gmx.net> wrote:
> >
> > >I always
> > >dissolve and dilute my primers in double distilled water and this
> > >worked always.
> >
> > One more thing: I always dilute the primer stocks to 100pmol and make
> > a 10pmol dilution for use in PCR.
> >
> > Christian
> >
> > --
> >        [X] <-- nail here for new monitor
> >
> >
> > ------------------------------
> >
> > Message: 12
> > Date: Tue, 28 Feb 2006 15:28:55 +0000
> > From: Christian Praetorius <prae at gmx.net>
> > Subject: Re: Primers, DDWater, TE Buffer
> > To: methods at net.bio.net
> > Message-ID: <46j8hkFb5bb5U3 at individual.net>
> > Content-Type: text/plain; charset=us-ascii
> >
> > "GysdeJongh" <jongh711 at planet.nl> wrote:
> >
> > >I  _think_  you mean 100 pmol / microliter
> >
> > Argh, you are right.
> >
> > Christian
> >
> > --
> >        [X] <-- nail here for new monitor
> >
> >
> > ------------------------------
> >
> > _______________________________________________
> > Methods mailing list
> > Methods at net.bio.net
> > http://www.bio.net/biomail/listinfo/methods
> >
> > End of Methods Digest, Vol 9, Issue 28
> > **************************************
> >
>
>
>--
>Sidharth Parida
>Senior Faculty
>Center for biotechnology and Research
>Neelachal Inst of Medical Sciences
>O.C.H.C building , Near Ram Mandir
>Bhubaneswar-3
>ORISSA
>09437089337(M)
>sidharth4 at gmail.com
>
>_______________________________________________
>Methods mailing list
>Methods at net.bio.net
>http://www.bio.net/biomail/listinfo/methods

***************************************
Dr. Eva Freisinger
University of Zurich
Institute of Inorganic Chemistry
Winterthurerstrasse 190, Room 34-F-44
CH-8057 Zurich
Switzerland

Phone: +41 (0)44 635 4621 (office)
        +41 (0)44 635 4660 (lab)
Fax:   +41 (0)44 635 6802
http://www.aci.unizh.ch/
***************************************



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