We report about the efficiency of the click reaction directly on PCR products containing alkyne-modified uridine nucleosides. Galactose azide was clicked onto single-stranded and double-stranded 300mer DNA in order to investigate the efficiency of sugar labeling of DNA under various conditions. Single-stranded alkyne-modified DNA was prepared by selective lambda exonuclease digestion of 5′-phosphate-labeled double strands. A comparison of the click reaction efficiency on modified single and double stranded 300mers under identical conditions shows a pronounced effect of the strandedness on the yield of the click reaction.

The efficiency of the click reaction on modified single- and double-stranded DNA is significantly affected by the strandedness of the DNA.

Pronounced Effect of DNA Hybridization on Click Reaction Efficiency

Key Takeaway: The efficiency of the click reaction on modified single- and double-stranded DNA is significantly affected by the strandedness of the DNA.