How to covalently link DNA to beads?

martin LEACH leach at bu.edu
Mon Feb 26 18:20:08 EST 1996


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)
: ---------------------------------------------------------



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