Hi there, I attempted to conjugate some short (~20bp) oligo probes using a commercial kit, but I believe the conjugation failed due to the use of an incorrect filter size. I still have the TCEP-reduced oligos, which I purified via ethanol precipitation, stored in water at 4C. Is it advisable to attempt the conjugation again with the reduced oligos if it has been over two weeks? Cheers
Hi,
I think that, while I do not have experience re-using deprotected DNA-SH after a few days, it wouldn't harm to add a small excess of TCEP to ensure that any thiol groups are reduced (they tend to form DNA-SS-DNA in solution and in the presence of oxygen) and attempt the conjugation again.
I am basing my answer in the fact that in a few occasions, doing reactions that involve DNA-SH, if I did not get the equivalents exactly right and there was a small excess of DNA-SH, I could still see the free DNA-SH in the LC-MS analysis 24 h later.
I hope that this helps.
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