Lepidine Orange derivative as a new dye for sensitive fluorescent detection of DNA

Yu V Didan,I Ya Dubey,D V Kryvorotenko,V V Negrutska

doi:10.7124/bc.000842

Abstract

The aim. To study new cyanine dye LO-7 as a reagent for dsDNA visualization in electrophoretic gels. Methods. Gel electrophoresis, restriction, fluorescence detection, mobility shift assay. Results. LO-7 binds to DNA to form stable highly fluorescent complexes. As little as 80 pg DNA can be detected in LO-7-stained agarose gel using a laser scanner, and 0.3 ng with UV-transilluminator. This sensitivity is several times higher than can be achieved with ethidium bromide, and close to that of SYBR Green I. Conclusions. LO-7 belongs to the most efficient stains for dsDNA visualization and thus can be used in bioanalytical applications where high sensitivity is required.

Highlights

Non-radioactive labeling of biomolecules has been studied extensively for several decades
The majority of fluorescent dyes commonly used for nucleic acids visualization assays belong to the cyanine family [2, 5], since these molecules display high affinity for nucleic acids, strong absorption in the visible spectral range and often sharp increase of fluorescence when interacting with nucleic acids [6,7,8]
We present a new cyanine dye LO-7 (Lepidine Orange derivative), a structural analog of well known dyes Thiazole Orange [9] and SYBR Green I from «Mo

Summary

Introduction

Non-radioactive labeling of biomolecules has been studied extensively for several decades. Ó Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013 lecular Probes» (USA) ( «Invitrogen») [10, 11], as a new highly sensitive fluorescent stain for DNA detection in agarose gels. Plasmid DNA was restricted with BstNI enzyme («Fermentas») and serially diluted (1:4) to get 4 samples with DNA concentrations of 23, 5.75, 1.44 and 0.36 ng/ml.

Results

Conclusion