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[Siddhartha Parida]

What is composition of electro transfer buffer for transferring DNA to
nitrocellulose membranes
by electroblotting  ?


On Tue, Mar 18, 2008 at 10:34 PM,  <methods-request from oat.bio.indiana.edu> wrote:
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>  Today's Topics:
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>    1. Re: "freeze and squeeze" DNA extraction (Farahani Poupak)
>    2. Re: Digestion Problem (Carla Beukes)
>    3. Re: DMSO in PCR (DK)
>    4. Re: DMSO in PCR (chovek69)
>
>
>  ----------------------------------------------------------------------
>
>  Message: 1
>  Date: Mon, 17 Mar 2008 10:07:01 -0700 (PDT)
>  From: Farahani Poupak <poupakfar from yahoo.com>
>  Subject: Re: "freeze and squeeze" DNA extraction
>  To: Sudheendra Rao N R <sudhee26 from gmail.com>,    "Dunowska, Magda"
>         <M.Dunowska from massey.ac.nz>,      methods <Methods from magpie.bio.indiana.edu>
>  Message-ID: <54812.85425.qm from web51610.mail.re2.yahoo.com>
>  Content-Type: text/plain; charset=iso-8859-1
>
>  I just use "Freeze 'N Squeeze DNA Gel Extraction Kit"
>  from Bio-Rad. I don't have any variants for this
>  protocol.
>
>
>
>
>  --- Sudheendra Rao N R <sudhee26 from gmail.com> wrote:
>
>  > I use this method.QIAEX II particle.
>  >
>  http://www1.qiagen.com/literature/handbooks/literature.aspx?id=1000202
>  >
>  > try.
>  > sudhee.
>  >
>  > On Sun, Mar 16, 2008 at 3:19 PM, Dunowska, Magda
>  > <M.Dunowska from massey.ac.nz>
>  > wrote:
>  >
>  > > Hi,
>  > >
>  > > Sorry to revisit the subject - I have just gone
>  > through the archives and
>  > > can't find a description of a method I used to use
>  > a few years ago to get
>  > > DNA from agarose gels. I used normal (not LMP)
>  > agarose and TBE buffer. All I
>  > > remember is that it involved freezing and thawing
>  > a gel slice, but I think I
>  > > was also adding water or TE (can't remember what
>  > and how much), heating the
>  > > thawed gel to either 65 or 70 degrees for 5 or 10
>  > minutes (?) and mashing it
>  > > all up with the blue tip before centrifuging
>  > everything at 4 degrees to
>  > > collect supernatant.
>  > >
>  > >
>  > >
>  > > I just got back to the molecular work after a few
>  > years in a slightly
>  > > different role, and (surprisingly...) can't recall
>  > all the details of
>  > > procedures that I used to do on a daily basis and
>  > never bothered to write
>  > > down outside of the lab books that are no longer
>  > with me...Anybody out there
>  > > who uses/used a similar variant of the "freeze and
>  > squeeze" and can give me
>  > > the details?
>  > >
>  > > Thanks!
>  > >
>  > > Magda
>  > >
>  > >
>  > >
>  > >
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>  ------------------------------
>
>  Message: 2
>  Date: Tue, 18 Mar 2008 07:49:49 +0200
>  From: "Carla Beukes" <carla.beukes from gmail.com>
>  Subject: Re: Digestion Problem
>  To: methods from magpie.bio.indiana.edu
>  Message-ID:
>         <cfd68bd0803172249s141eabbfkdd31e920fdc2747d from mail.gmail.com>
>  Content-Type: text/plain; charset=ISO-8859-1
>
>  Thank you so much.  Will certainly try that.
>
>  Carla
>
>
>  ------------------------------
>
>  Message: 3
>  Date: Mon, 17 Mar 2008 23:03:30 GMT
>  From: dk from no.email.thankstospam.net (DK)
>  Subject: Re: DMSO in PCR
>  To: methods from net.bio.net
>  Message-ID: <7NCDj.51$XB6.34 from newsfe06.lga>
>
>  In article <mailman.1120.1205773333.2451.methods from net.bio.net>, "popi Arapini" <karapini from gmail.com> wrote:
>  >hI! everybody,
>  >I'am using DMSO to standardize my PCR (high GC content)
>  >I've read in a protocol that, once DMSO in the thermowell tube, it shall
>  >not  be spinned  down . Does anybody  know why??
>
>  No reason. You heard wrong.
>
>
>
>  ------------------------------
>
>  Message: 4
>  Date: Tue, 18 Mar 2008 03:46:08 -0700 (PDT)
>  From: chovek69 <ivanoov from gmail.com>
>  Subject: Re: DMSO in PCR
>  To: methods from net.bio.net
>  Message-ID:
>         <e318808a-a1e6-432d-a409-a4a887fd4937 from m34g2000hsc.googlegroups.com>
>  Content-Type: text/plain; charset=ISO-8859-1
>
>  On Mar 18, 1:03 am, d... from no.email.thankstospam.net (DK) wrote:
>  > In article <mailman.1120.1205773333.2451.meth... from net.bio.net>, "popi Arapini" <karap... from gmail.com> wrote:
>  >
>  > >hI! everybody,
>  > >I'am using DMSO to standardize my PCR (high GC content)
>  > >I've read in a protocol that, once DMSO in the thermowell tube, it shall
>  > >not  be spinned  down . Does anybody  know why??
>  >
>  > No reason. You heard wrong.
>
>  Yes this is nonsense. 5-10% DMSO disolves perfectly in PCR buffer ....
>  so no bother. In our lab 5% DMSO seems best for many assays.
>
>
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>  End of Methods Digest, Vol 34, Issue 18
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>



-- 
Sidharth Parida
Senior Faculty
Center for biotechnology and Research
Neelachal Inst of Medical Sciences
O.C.H.C Complex, Block - III (Near Ram Mandir)
Janpath,Unit-3,Bhubaneswar- 751001
ORISSA
09437089337(M)

sidharth4 from gmail.com
www.nimsindia.com