Mircury lna universal rt

Manufactured by Qiagen

The MiRCURY LNA Universal RT is a lab equipment product designed for reverse transcription (RT) of microRNA (miRNA) and other small RNA molecules. It provides a universal solution for converting miRNA and small RNA into cDNA, which can then be used for downstream analysis and applications.

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Lab products found in correlation

Lightcycler 480, by Roche (1 mentions) Sybr green master mix, by Roche (1 mentions) Lightcycler 480 machine, by Roche (1 mentions) Lna primers, by Qiagen (1 mentions) Taqman reagent, by Thermo Fisher Scientific (1 mentions) Sybr qpcr master mix, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Mirneasy mini kit, by Qiagen (1 mentions) Quantitect rt kit, by Qiagen (1 mentions)

Close spelling variants of this product

MiRCURY LNATM Universal RT microRNA PCR kit (13 citations) MiRCURY LNATM Universal RT microRNA PCR system (6 citations) MiRCURY LNATM Universal RT microRNA PCR (5 citations) LNA miRCuRY (2 citations)

3 protocols using mircury lna universal rt

Quantification of miR-34 Family Members

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Fifty ng of RNA was reverse transcribed (RT) using miRCURY LNA™ Universal RT (Exiqon). RT-qPCR for miR34A-5p, miR34A-3p, miR34B-5p, miR34B-3p, miR34C-5p and miR34C-3p was performed using miRCURY LNA™ Universal RT primer sets (nomenclature according to miRbase18), and U44 RNA was used for normalization (prod. No. 204486, 204318, 204005, 204424, 204373, 204407, 203902, respectively). The RT-qPCRs were performed on the LightCycler 480 instrument (Roche diagnostics) using the conditions recommended by Exiqon. Absolute quantification was performed using a miR-34A-5p mimic in final amounts of 1x10⁸ copies to 10 copies per RT reaction.

Asmar F., Hother C., Kulosman G., Treppendahl M.B., Nielsen H.M., Ralfkiaer U., Pedersen A., Møller M.B., Ralfkiaer E., de Nully Brown P, & Grønbæk K. (2014). Diffuse large B-cell lymphoma with combined TP53 mutation and MIR34A methylation: Another “double hit” lymphoma with very poor outcome?. Oncotarget, 5(7), 1912-1925.

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Quantification of miRNA Expression in OPMD

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RNA was converted to cDNA in the presence of spikes, sp3 and sp6, using the Exiqon’s miRCURY LNA Universal RT. Pipetting was carried out with a robot; PCR amplification was carried out with miRNAs specific primers and EXILENT Sybr green master mix in technical duplicates in a Roche light cycler 480 machine. CT values were made from the average of duplicates. CT > 38 was excluded. Expression levels were normalized to the average of sp3 and sp6, using the dCT calculation. In saliva, normalization was also to miR23a, since its product was amplified in all samples, and its levels were unchanged between control and OPMD. In serum, this step was omitted, since PCR product was not detected in all samples. When PCR products were obtained in only half of the control subjects or OPMD samples, it was excluded from the analysis.

Raz V., Kroon R.H., Mei H., Riaz M., Buermans H., Lassche S., Horlings C., Swart B.D., Kalf J., Harish P., Vissing J., Kielbasa S, & van Engelen B.G. (2020). Age-Associated Salivary MicroRNA Biomarkers for Oculopharyngeal Muscular Dystrophy. International Journal of Molecular Sciences, 21(17), 6059.

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Gene and MicroRNA Expression Analysis

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RNA was extracted using RNeasy Mini kit (Qiagen) or by using the TRIzol method (Qiagen). qPCR was carried out using the QuantiTect RT kit (Qiagen) according to the manufacturer’s protocol. qPCR was carried out using the ViiA7 qPCR system with TaqMan reagents (Life Technologies). Gene expression levels were normalized to HPRT for all cell types.
Total RNA for miRNA analysis was extracted with miRNeasy Mini kit (Qiagen) or by using the Trizol method. miRNA-RT was carried out using miRCURY LNA Universal RT according to the manufacturer’s instructions (Exiqon), and SYBR qPCR master mix (Life Technologies) was used for quantitative PCR. Three LNA primers (Exiqon) designed to detect the most common miR-499a-5p isomiRs (miRBase) were pooled for detection. The miR-208b LNA primer was optimized by using a primer with a lower Tm. SNORD38B was used as an endogenous control.

Jarret A., McFarland A.P., Horner S.M., Kell A., Schwerk J., Hong M., Badil S., Joslyn R.C., Baker D.P., Carrington M., Hagedorn C.H., Gale M J.r, & Savan R. (2016). Hepatitis-C-virus-induced microRNAs dampen interferon-mediated antiviral signaling. Nature medicine, 22(12), 1475-1481.

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