LMP agarose gels run in TAE buffer (used to be:Purification from Low Melt)

Basavaraju Shankarappa bsh at MED.PITT.EDU
Mon Feb 28 13:41:09 EST 1994


>cle <1994Feb24.141630.22721 at alw.nih.gov> Jim Owens <jow at helix.nih.gov> writes:
> >From: Jim Owens <jow at helix.nih.gov>
> >Subject: Re: Purification from Low Melt
> >Date: Thu, 24 Feb 1994 14:16:30 GMT
> >In article <1994Feb18.180245.22274 at alw.nih.gov> Jim Owens,
> >jow at helix.nih.gov writes:
> >>before they get me.  Thus, my method involves using phenol and
> >chloroform.
> >>
> >>Cut the bands out and put in 1.5ml microfuge tubes.  Weigh the agarose in
> >>........blah, blah, blah left out......
> >From some e-mail responses I've gotten to my posting here, I should
> >mention one other detail about my protocol:
> >Recovery is decent only for LMP agarose gels run in TAE buffer.
> Jim and and other netters,
> 
> This is something that I have always done, but not known the reason why it is 
> so.  Could you please enlighten me as to the reason one should prepare LMP 
>agarose gels in TAE instead of TBE when purifying DNA fragments from the gel.  
> (I also always use TAE gels for Southern blotting to nylon membranes).
> | Dr. Vernon E. Coyne,                                               |
> | Microbiology Department,            Tel: (27-21) 650-3259/70       |
> | University of Cape Town,            Fax: (27-21) 650-4023          |
> | Private Bag,                        E-mail: vernon at micro.uct.ac.za |
> | Rondebosch, 7700                                                   |
> | South Africa                                                       |
> +--------------------------------------------------------------------+
> 
	I have been told that this is because the borate in TBE binds to
nucleic acid and thus it changes its properties.  If you remember the 
old geneclean kits, you could use it only on TBE gels until they came 
up with a TBE modifier so that it can be used on TBE gels also..
Raj Shankarappa
bsh at med.pitt.edu




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