So we got 3 prime thiol modified 14 mer oligonucleotides that we want to attach to FITC for detection via agarose gel electrophoresis.
I was wondering if I can still purify the DNA via spin column even though it has FITC by thiol modification in the 3' end.
If its possible, what spin column would you recommend?
I don't know which brand spin column you are using but, Fluorescein isothiocyanate, being an aromatic moiety should increase its affinity towards the column matrix, which means you might need to increase polarity of your elution solvent.
It also depends if your oligos have DMT-ON or OFF. Are you synthesizing oligos yourself?
Maybe i am completely wrong as I have not labeled oligos before, but instead peptides, we always used HPLC with a UV detector and fluorescence detector that way we could separate the labelled and unlabelled peptides. But maybe you don't care about the probably small portion of unlabelled oligos as they probably don't matter in the gel. So what I am saying is I don't think that the spin column will necessarily be able to separate the labelled from unlabelled oligo, if that is important to you.
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