Loading Combs for Sequencers

Skip Vaught, Catalina Apostol, Brian Coullahan, and Mark Skip Vaught, Catalina Apostol, Brian Coullahan, and Mark
Wed Jun 18 13:13:51 EST 1997


>Hi Everybody
>
>I posted a query at the end of last month about the use of a porous comb
>for the rapid (and potentially very easy and painless) loading of
>samples onto sequencing gels.
>
>The relevant article from Ansorge's group at the EMBL, Heidelberg, was
>published in Nucleic Acids Research (volume 25 (issue 11) pages 2229 -
>2230, year 1997).  All that is said that the material for the comb was
>made from MV Cellulose-Mischester from Macherey-Nagel or Nylon membrane
>from Boehringer Mannheim.
>
>Does anybody know the catalogue numbers for these membranes, their
>cost?  Can the combs be re-used?  Will they need washing with acid or
>alkali to regenerate a usable comb?!  In particular has anybody else
>tried this method?
>
>I have emailed Ansorge, but I am awaiting his reply........
>
>The Sanger Centre WEB pages (http://www.sanger.ac.uk/ and link to the
>poeple and hence to the development group) have a page on loading with a
>comb, but the link doesn't work yet, as they have just updated their WEB
>site.
>
>As this development seems to be extremely relevant for automated
>sequencing, I am amazed at the lack of discussion on the newsgroup, or
>is everybody too busy using the combs to be able to read the newsgroup
>and reply :-)
>
>The ability to load a comb on the bench and then insert it ontop of the
>gel seems to be a big adavantage to loading a gel in the apparatus with
>flat tips and trying not to spill over into the next lane, while at the
>same time explaining to somebody enquiring about their sequence run to
>go away as what I am doin gis extremely delicate and requires a lot of
>concentration.
>
>Mick
>--
>_____________________________________________________________________
>Dr M D Jones                      Tel: 081-383-3328 (+44-81-383-3328)
>Department of Virology          FAX: 081-383-2299 (+44-81-383-2299)
>RPMS,  Du Cane Road              Email: mjones at rpms.ac.uk
>London,  W12 0NN,  UK
>URL: http://www.rpms.ac.uk/departments/virology/virolhome.html
>***********************************************************
>"To Infinity and Beyond."   Buzz Lightyear (Toy Story)
>"Crackin' toast, Gromit."    Wallace (The Wrong Trousers)
>_____________________________________________________________________

Dear Dr. Mick,

The porous comb sounds like a great idea, but I don't think it will work
well for those of us using version 2 of ABI's analysis program. Because
lane tracking is from the bottom - up, we are required to load lanes in a
staggered fashion (odd numbered lanes- electrophorese- even numbered
lanes).

The comb might work on version 3 (it tracks from the middle - out), but
tracking was so unreliable, we switched back to Ver. 2.

Skip Vaught
DNA Sequencing Service
University of Arizona





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