Isothermal chemiluminescent assay based on circular stand-displacement polymerization reaction amplification for cel-miRNA-39-3p determination in cell extracts

Comparison of chemiluminescent heterogeneous and homogeneous–heterogeneous assays coupled with isothermal circular strand-displacement polymerization reaction amplification for the quantification of miRNA-141

“…The concentrations of the equimolar mixture of dNTPs (10 μM), primer (100 nM) and lKF (8 U μL −1 ) were the same as used in our previous work. 31…”

“…To estimate the yield of miRNA purification, a known quantity of miRNA-39 was added in the studied lysates and then after miRNA purification, its concentration was determined in the resulting solution by the assay reported by us recently. 31 In our work, miRNA-39 was chosen as a reference analyte because the miRNA of this nematode ( Caenorhabditis elegans ) is not present in human samples.…”

“…The concentrations of the equimolar mixture of dNTPs (10 µM), primer (100 nM) and lKF (8 U μL −1 ) were the same as used in our previous work. 31 As mentioned above, the proposed assay was developed using not only target-dependent amplification but also amplification of the detection system. The conditions for the performance of the enhanced chemiluminescence reaction have been studied previously in detail; 25 therefore in this work, we optimized only the concentration of the Str-PolyHRP80 conjugate (Fig.…”

Quantitation of MicroRNA-155 in Human Cells by Heterogeneous Enzyme-Linked Oligonucleotide Assay Coupled with Mismatched Catalytic Hairpin Assembly Reaction

“…The human cells (MCF-7, HeLa, Caco2, and HepG2) were obtained from the Russian Collection of Cell Cultures (Institute of Cytology, St. Petersburg, Russia). All cells were cultured in Dulbecco’s Modified Eagle’s Medium with 10% fetal bovine serum at 37 °C in an atmosphere containing 5% CO 2 as described in our previous work [ 19 ].…”

“…The purification of short RNAs from cell lysates was carried out using the LRU-100-50 kit from “BioLabMix” (Russia, Novosibirsk) as described previously [ 19 ]. Purified miRNA samples were stored at −20 °C.…”