Hi Yvonne, some possibly very weird ideas (sorry, I'm a chemist):
a) any chance to have a carboxylic group attached to the oligo? Then activate the carboxylic group by a carbodiimide method and couple to lysine residues of AP b) any chance to have one ribobase incorporated into your oligo? Then do periodate oxidation and link to lysines of AP, as described in standard protocols for HRP-labeling (it's just inverse, this time) c) any chance to get streptavidin attached to AP? then buy biotinylated oligos, mix and you're done. In worst case, just try to couple streptavidin and AP with a bifunctional cross-linker. d) as thiol modified oligos seem to be available (http:// www.metabion.com/products/dna_modifications.php), use a maleimide/ succinimide bifunctional linker to attach it to AP: Maleimide activated carboxyls react with SH groups, succinimide activated carboxyls with aliphatic amino groups. Thus, first react the oligo with the maleimide part. If desired, purify by gel filtration (either absolutely water-free or in 1mM HCl), then react with AP in 100mM Borate or Carbonate, pH8-9. Tris or similar aminogroup compounds as well as BME, DTT etc are absolutely to avoid, of course, they'll screw up the couplings. Hopefully, these thoughts will lead you to something feasible... /Wo On Sep 23, 4:29 pm, "Yvonne Couch" <yvonne.co...@pharm.ox.ac.uk> wrote: > Hi all, > > I'm in the UK and am looking for a company that might sell alkaline > phosphatase labelled oligonucleotides for in situ hybridization - anyone > have any ideas? > > Thanks > > Yvonne _______________________________________________ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
