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. 1993 Aug 11;21(16):3737–3743. doi: 10.1093/nar/21.16.3737

Tritium labeling of antisense oligonucleotides by exchange with tritiated water.

M J Graham 1, S M Freier 1, R M Crooke 1, D J Ecker 1, R N Maslova 1, E A Lesnik 1
PMCID: PMC309879  PMID: 8367289

Abstract

We describe a simple, efficient, procedure for labeling oligonucleotides to high specific activity (< 1 x 10(8) cpm/mumol) by hydrogen exchange with tritiated water at the C8 positions of purines in the presence of beta-mercaptoethanol, an effective radical scavenger. Approximately 90% of the starting material is recovered as intact, labeled oligonucleotide. The radiolabeled compounds are stable in biological systems; greater than 90% of the specific activity is retained after 72 hr incubation at 37 degrees C in serum-containing media. Data obtained from in vitro cellular uptake experiments using oligonucleotides labeled by this method are similar to those obtained using 35S or 14C-labeled compounds. Because this protocol is solely dependent upon the existence of purine residues, it should be useful for radiolabeling modified as well as unmodified phosphodiester oligonucleotides.

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Selected References

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