Journal
ANALYTICAL SCIENCES
Volume 29, Issue 6, Pages 605-610Publisher
JAPAN SOC ANALYTICAL CHEMISTRY
DOI: 10.2116/analsci.29.605
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Funding
- Natural Science Foundation of China [21025521, 20875027]
- National Key Basic Research Program [2011CB911000]
- CSIRT Program
- Natural Science Foundation of Hunan Province [10JJ7002]
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Considering the crucial role played by microRNAs (miRNAs) in biological processes, we developed a novel strategy for simple and colorimetric detection of miRNA by combining target amplification with DNAzyme. Throughout the work, a 22-nt oligonucleotide sequence was used as a model analyte. A label-free hairpin probe (HP) was used as a simple platform for sensing the target. In the presence of the target, the HP was opened, and then the isothermal circular strand-displacement process occurred with the help of a primer, deoxynucleotide solution mixture (dNTPs), Klenow fragment exo(-) polymerase, and Nb.BbvCI nicking enzyme. As a result, the target was recycled and multicopies of target analogues were generated that function in the same manner as the target, accompanied by the accumulation of signal elements. In this work, as low as 0.5 fM nucleic acid target was detected by horseradish peroxidase-mimicking DNAzyme catalyzing the oxidation of ABTS(2-) to colored ABTS(.-).
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