Electrochemical detection of DNase I activity

Abstract

DNase I in one microl of the water could quantitate electrochemically with the detection limit of 0.01 units (ca. 20 pg) by using the ferrocenyl oligonucleotide-immobilized electrode prepared by thiolated oligonucleotide and ferrocenyl carbodiimide as a simple labeling reagent of redox unit.

Ferrocenylnaphthalene Diimide-based Electrochemical Ribonuclease Assay

“…This behavior is similar to previous work, where electrochemically active DNA ligand showed a lower slope than that after hybridization with the complementary DNA. 15 The ligand was more concentrated to the doublestranded DNA than to the single-stranded DNA on the electrode. This also showed that 1 could be used for estimating the amount of mRNA on the electrode, and could detect a decreasing amount of mRNA on the electrode after digestion with RNase.…”

“…[7][8][9] Electrochemistry is generally expected to achieve high-sensitive analysis with an inexpensive, compact instrument; this technique has been applied to DNA detection. [7][8][9][10][11][12][13][14][15][16][17][18][19][20][21] Electrochemical DNA analysis so far reported is briefly grouped into two methods based on the ways to be used for electrochemically active labeling of DNA [10][11][12][13][14][15] and an electrochemically active DNA indicator. [7][8][9][16][17][18][19][20][21] Highly sensitive DNA detection was achieved in both methods.…”

“…[7][8][9][16][17][18][19][20][21] Highly sensitive DNA detection was achieved in both methods. An electrochemical DNA enzyme assay has also been reported in DNA polymerase, 14 telomerase, 8 DNase I, 15 and S1 nuclease. 13 However, an electrochemical RNase assay has not yet been reported to the best of our knowledge.…”

Universal Platform for Sensitive and Label-Free Nuclease Assay Based on Conjugated Polymer and DNA/Intercalating Dye Complex

“…DNA or RNA cleavage reactions catalyzed by nucleases such as restriction nucleases and nonspecific nucleases are essential in a variety of fields ranging from biotechnology to pharmacology, as well as in biological processes involving replication, recombination, DNA repair, molecular cloning, genotyping, and mapping. − Recent attempts to attach restriction endonucleases to nanostructures have been undertaken due to the well-established ability of the endonucleases to cleave DNA at specific recognition sites. − Traditional techniques, including gel electrophoresis, high-performance liquid chromatography (HPLC), electrochemical study, and enzyme-linked immunosorbent assay (ELISA) have been established in assaying nuclease activities. − These protocols share the drawbacks of being time-intensive, DNA-consuming, discontinuous, laborious, and usually requiring isotope labeling. Many of these limitations are now being addressed by the development of fluorescence assays based on fluorescence quenching or fluorescence resonance energy transfer (FRET). − Although promising, these techniques are compromised by the requirement for double-labeled DNA probes, limited chemical stability, and interferences by external nonspecific events.…”