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Updated: May 24, 2026

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
Published on: July 8, 2025
Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis
Asfia Sultana1, Anupriya M Geethakumari1, Kabir H Biswas1
1College of Health and Life Sciences, Hamad Bin Khalifa University, Doha, Qatar.
Abstract:
NanoLuc (NLuc) has been conjugated with DNA oligonucleotides (DNA oligo) for biosensing applications, which requires assessment of bioluminescence activity and conjugation efficiency. Here, we describe a straightforward, non-denaturing agarose gel electrophoresis-based assay that enables simultaneous monitoring of NLuc bioluminescence activity and NLuc-DNA oligo conjugation. Agarose gel electrophoresis of NLuc-DNA conjugation reaction samples and bioluminescence imaging revealed NLuc bioluminescence activity and a mobility shift of the protein confirming conjugation. Further, quantitative bioluminescence imaging allowed determination of active NLuc fraction and NLuc-DNA conjugation efficiency. The method reported here could be broadly applicable to other bioluminescent protein-DNA oligo conjugation workflows.

