How to covalently link DNA to beads?

martin LEACH leach at bu.edu
Tue Feb 27 10:51:50 EST 1996


Well, 

according to the chemists at ABI/perkin elmer...there is no way of
deprotecting without cleavage..oh well could have worked...

perhaps it may still work with chemistries other than carruthers & beaucage
(used on the abi machines)

Martin

martin LEACH (leach at bu.edu) wrote:
: On a similar note....using a DNA synthesizer create your custom oligo...the
: oligo will remain bound to the glass beads they are synthesized (unless you
: hydrolyze the siloxane bond with NH4OH)....you want the trityl groups removed
: during synthesis. Then when finished dismantle the synthesis column and
: harvest
: the glass beads with oligo still attached.

: only problem is that this DNA is still in a form as base-protected
: phosphotriesters...(can you tell I am reading from the ABI manual..;)...they
: are usually deprotected using the NH4OH...but this also cleaves them from the
: column.....if there was a way of deprotecting without cleavage (a weaker
: base?? instead of NH4OH) then perhaps you could create your desired oligo this
: way??

: just my thruppeny bit...

: Martin



: : Marc:

: : How about this?  Make custom oligos (with attached 5' biotin or other "handle") that will anneal with the lambda cos site.  Mix with=
: :  your lambda DNA, link with ligase, then attach to streptavidin beads.

: : - Peter

: : ---------------------------------------------------------
: : Peter Wang, M.D., Ph.D.
: : MRC Centre for Protein Engineering,
: : Hills Road, Cambridge, CB2 2QH, England

: : Tel (01223) 402104  (international calls +44-1223-402104)
: : Fax (01223) 402140  (     "          "   +44-1223-402140)
: : ---------------------------------------------------------



: --
: .....          Martin Leach                Email:leach at bu.edu
:    _|____      Dept. of Pharmacology       Phone: (617) 638-5323
:    / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329
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: My homepage: http://155.41.115.114/Leach.html
: Biotech company WWW REgistry has moved to: http://www.data-transport.com
: Check out the Biotech Rumor mill: http://155.41.115.114/jumper/rumor.html

--
.....          Martin Leach                Email:leach at bu.edu
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329
 _/  |-/__==/  80 E. Concord St. (L603)
(BULLDOZER) \_ Boston MA 02118            "Not the old underpants
               USA                                on your
                                             head.....WIBBLE"

My homepage: http://155.41.115.114/Leach.html
Biotech company WWW REgistry has moved to: http://www.data-transport.com
Check out the Biotech Rumor mill: http://155.41.115.114/jumper/rumor.html



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