I had purified a fluorescent labeled oligo through HPLC. I had collected the peaks from different runs and pooled them and lyophilized. When I run it again on HPLC it is giving multiple peaks. What could be the reason? Does the process of lyophilization degrade or break the bond between the fluorophore and the oligo.
It sounds like the fluorescent dye has separated from the oligo. That is usually the course of the two peaks. Normally, this does not happen in freeze drying or centrifugal evaporation provided that the samples are protected from light, particularly UV and also from heat.
Kind regards, Rob
Thanks Rob.
But there are many peaks, not just two. The samples were properly protected and kept under chilled conditions at all times. I think there could be something more to this degradation.
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