Mircury lnatm universal rt microrna pcr

Manufactured by Qiagen

Sourced in Germany

The MiRCURY LNATM Universal RT microRNA PCR is a laboratory instrument designed for the detection and quantification of microRNA (miRNA) expression. It utilizes a universal reverse transcription (RT) reaction and a microRNA-specific quantitative PCR (qPCR) approach to enable sensitive and accurate miRNA profiling.

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Lab products found in correlation

Exilent sybr green master mix, by Qiagen (2 mentions) Universal cdna synthesis kit 2, by Qiagen (2 mentions) Ms2 rna, by Roche (1 mentions) Lightcycler 480, by Roche (1 mentions) Abi 7500 instrument, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by Qiagen (1 mentions)

Close spelling variants of this product

MiRCURY LNA™ Universal RT microRNA PCR MiRCURY LNATM Universal RT microRNA PCR kit MiRCURY LNATM Universal RT microRNA PCR system MiRCURY LNA Universal RT microRNA MiRCURY LNA microRNA PCR MiRCURY LNA Universal RT LNA miRCuRY

5 protocols using mircury lnatm universal rt microrna pcr

Quantifying miR-9 Expression in Carcinoma Cells

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cDNA was synthesized using miRCURY LNA^TM Universal RT microRNA PCR, Universal cDNA Synthesis Kit II (Qiagen), according to the manufacturer’s instructions.
The expression level of miR-9 was obtained using miRCURY LNA^TM Universal RT microRNA PCR, Exilent SYBR^® Green master mix (Qiagen). Quantitative PCR (qPCR) amplifications were performed in triplicate using the primers for hsa-miR-9-5p (YP00204513 - Qiagen) and hsa-SNORD49A (Qiagen) [44 (link)] on a LightCycler 480 (Roche Berlin, Germany). The quantification cycle (Cq) was used to calculate the relative miR-9 expression using normalization to hsa-SNORD49A (ΔCq).
miR-9 expression was compared between patients from the Low and High lamin A group, or between EMA− and EMA+ carcinoma cells, corresponding to lamin A− and lamin A+ carcinoma cells, respectively (ΔΔCq). miR-9 relative expression (fold change (Fc)) was calculated by the 2^−ΔΔCq method [36 (link)]. miR-9 was considered as upregulated if the fold change was ≥ 2 and down-regulated if the fold change was ≤ 0.5.

Guinde J., Benoit A., Frankel D., Robert S., Ostacolo K., Lévy N., Astoul P., Roll P, & Kaspi E. (2020). miR-9 Does Not Regulate Lamin A Expression in Metastatic Cells from Lung Adenocarcinoma. International Journal of Molecular Sciences, 21(5), 1599.

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Serum miRNA Isolation and Analysis

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Serum miRNAs were isolated from serum with DANAGENE microRNA and Cell free RNA Kit (DANAGEN, Badalona, Spain) using MS2 RNA (Roche, Basel, Switzerland) as a carrier to improve the RNA isolation and exogenous controls (Unisp2,4 and 5). miRNAs were retrotranscribed into a cDNA with the miRCURY LNA^TM Universal RT microRNA PCR (EXIQON).

Carrasco‐Rozas A., Fernández‐Simón E., Lleixà M.C., Belmonte I., Pedrosa-Hernandez I., Montiel-Morillo E., Nuñez‐Peralta C., Llauger Rossello J., Segovia S., De Luna N., Suarez‐Calvet X., Illa I., Díaz‐Manera J, & Gallardo E. (2019). Identification of serum microRNAs as potential biomarkers in Pompe disease. Annals of Clinical and Translational Neurology, 6(7), 1214-1224.

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Quantification of microRNA expression

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Each sample of 20 ng RNA was reverse transcribed using a miRCURY LNA^TM Universal RT microRNA PCR (203907, Exiqon, Vedbaek, Copenhagen). Quantification of specific miRNAs relative expression was carried out with a miRCURY LNA microRNA PCR, ExiLENT SYBR Green master mix (203401, Exiqon, Vedbaek, Copenhagen). For the PCR reaction, cDNA was diluted 60 times. As a reference, hsa-miR-103a-3p was used. Primers for hsa-miR-324-3p (YP00204303), hsa-miR-328-3p (YP00204364), hsa-miR-99b-5p (YP00205983), hsa-miR-1260a (YP00205892), hsa-miR-361-5p (YP00206054), hsa-miR-484 (YP00205636), and the reference hsa-miR-103a-3p (YP00204063) were provided by Exiqon. All experiments were carried out three times. For the relative quantification of miRNA levels, the ‘ΔΔCq’ method was used [33 (link)].

Nowak I., Boratyn E., Student S., Bernhart S.F., Fallmann J., Durbas M., Stadler P.F, & Rokita H. (2020). MCPIP1 ribonuclease can bind and cleave AURKA mRNA in MYCN-amplified neuroblastoma cells. RNA Biology, 18(1), 144-156.

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Quantitative Analysis of Extracellular miRNAs

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Total Isolation and Quantitative Real-Time PCR miRNAs Analysis Total RNA was isolated from MVs using TRIzol Reagent, according to the manufacturer's instructions and measured by absorbance at 260 nm with a NanoDrop 1000 Spectrophotometer. RNA was reverse transcribed into cDNA using Universal cDNA synthesis kit II (Exiqon). The resulting cDNA transcript were used for PCR ampli cation using ExiLENT SYBR® Green Master Mix II (Exiqon) and miRNA speci c primer set (miRCURY LNA TM Universal RT microRNA PCR, Exiqon) for miR-223-3p, miR-181a-5p, miR-146a-5p, miR-125a-5p, miR-30c-5p and miR-23a-3p. The relative miRNA levels were calculated using the comparative Ct method, using miR-16-5p reference gene as endogenous control. miRNAs were chosen accordingly to previously published works in our research team (13) .

D’Amico E., Allegretta C., Zanghì A., Manuti V., Amoruso A., & avolio c. (2022). MicroRNAs 181a and 125a are highly expressed in early diagnosed RRMS: a pilot case-control study.

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Quantification of miR-210 Expression

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For miR-210 analysis, cDNA was obtained by reverse transcription with miRCURY LNA^TM Universal RT microRNA PCR (Exiqon-Qiagen, Hilden, Germany). Final reaction volumes were 10 µL containing 2 µL of reaction buffer, 1 μL Reverse transcription mix, 5 µL of RNase-free water and 2 µL of template RNA. Real-time miRNA detection was performed using miRCURY LNA^TM Universal RT microRNA PCR ExiLENT SYBR^® Green (Exiqon-Qiagen, Hilden, Germany) with 10 µL mixtures containing 5 µL of SYBR Green PCR Master mix, 1 of Primer Assay, and 4 µL of cDNA template. To normalize the miRNA expression, U6 snRNA (small nuclear RNA) expression was also quantified. The parameters for PCR amplification were 95 °C for 10 min followed by 40 cycles of 95 °C for 10 s and 60 °C for 1 min. miRNA qPCR was performed on ABI 7500 instrument (Applied Biosystems, Carlsbad, CA, USA). Each reaction was run in triplicate with a non-template control. The relative expression was calculated by using the comparative delta Ct method. Data were expressed as fold-change relative to the mean of U6 values.

Cristofaro I., Limongi C., Piscopo P., Crestini A., Guerriero C., Fiore M., Conti L., Confaloni A, & Tata A.M. (2020). M2 Receptor Activation Counteracts the Glioblastoma Cancer Stem Cell Response to Hypoxia Condition. International Journal of Molecular Sciences, 21(5), 1700.

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