Mircury lna microrna array power labeling kit

Manufactured by Qiagen

Sourced in Denmark

The MiRCURY LNA microRNA Array Power labeling kit is a laboratory equipment product designed for the labeling of microRNA samples prior to their detection and analysis using microarray technology. The kit provides the necessary reagents and protocols to label microRNA samples in a standardized and efficient manner, enabling accurate and reliable microRNA expression profiling.

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Lab products found in correlation

Hs 400 pro hybridization station, by Tecan (2 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Rnu6b, by Thermo Fisher Scientific (1 mentions) Genepix pro software, by Molecular Devices (1 mentions) Mircury lna microrna arrays mirbase v 10, by Qiagen (1 mentions) Genepix 4100a microarray scanner, by Molecular Devices (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Jmp pro 11, by SAS Institute (1 mentions) 3d gene human mirna oligo chip ver 17.0, by Toray (1 mentions) 3d gene rna extraction reagent, by Toray (1 mentions) Command console software v 4, by Thermo Fisher Scientific (1 mentions) Genechip scanner 3000 system, by Thermo Fisher Scientific (1 mentions) Mircury lna microrna chips, by Qiagen (1 mentions) Genepix pro 6, by Molecular Devices (1 mentions) Cy5 mono reactive dye, by GE Healthcare (1 mentions)

7 protocols using mircury lna microrna array power labeling kit

Murine microRNA Expression Profiling

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Hy3-/Hy-5 labelled RNA (miRCURY LNA microRNA Array Power Labeling Kit, Exiqon, Vedbaek, Germany) was manually hybridized on miRNA microarrays (miRCURY^TM LNA microRNA Array v10, Exiqon), which comprised 580 murine miRNAs (miRBase release 10.0). Slides were scanned by a GenePix 4000A Microarray Scanner (Axon Instruments, Foster City, USA) and analyzed by GenePix^® Pro Software (Axon Instruments). Differential expression of miRNAs was identified using a linear model approach (limma, Bioconductor analysis suite57 (link)). After background subtraction, non-positive spots were removed and the remaining signal intensity values were normalized to a small control RNA (RNU6B) (Life technologies, Carlsbad, USA). P-values were adjusted to multiple testing by Bonferroni correction.

Bartel S., Schulz N., Alessandrini F., Schamberger A.C., Pagel P., Theis F.J., Milger K., Noessner E., Stick S.M., Kicic A., Eickelberg O., Freishtat R.J, & Krauss-Etschmann S. (2017). Pulmonary microRNA profiles identify involvement of Creb1 and Sec14l3 in bronchial epithelial changes in allergic asthma. Scientific Reports, 7, 46026.

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miRNA Profiling Using LNA Microarrays

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Total RNA was extracted by using TRIzol (Invitrogen) according to the manufacturer’s instructions. 2.5 μg of total RNA were labeled with either Hy3 or Hy5, following the manufacture’s protocol (miRCURY LNA microRNA Array Power labeling kit, Exiqon). Two-color hybridization was carried out at 53°C for 16 hours using miRCURY LNA microRNA Arrays (miRBase v.10.0, Exiqon). Hybridization and washing steps were performed on the HS 400 PRO hybridization station (Tecan). The signal intensities were acquired by scanning the arrays on the GenePix 4100A microarray scanner (Molecular Devices, GenePix Pro 6.0.1.25 software). The obtained data were analyzed by using the Limma package (Smyth, G.K. 2005) from the Bioconductor Project (Bioconductor 2.5 on R 2.10), performing background correction (normexp, cutoff=10) and within–array normalization (global LOESS) as well as between-array normalization (scale method). Differential expression and statistical significance were assessed by applying linear model fit and empirical Bayes method (lmFit, eBayes). The default table of Limma is ranked by B-statistics (B), closely related to the adjusted p-value. The B-statistic used for this analysis is the log-odds that that gene is differentially expressed and it is automatically adjusted for multiple testing by assuming that 1% of the genes are expected to be differentially expressed.

Caporali A., Meloni M., Nailor A., Mitić T., Shantikumar S., Riu F., Sala-Newby G.B., Rose L., Besnier M., Katare R., Voellenkle C., Verkade P., Martelli F., Madeddu P, & Emanueli C. (2015). p75NTR-dependent activation of NF-κB regulates microRNA-503 transcription and pericyte–endothelial crosstalk in diabetes after limb ischaemia. Nature Communications, 6, 8024.

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Plasma miRNA Profiling in sALS

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Blood was collected in disodium ethylenediaminotetraacetate (Na2EDTA) tubes and centrifuged immediately, after which the plasma was separated and frozen at −80 °C. RNA (total RNA) was extracted from 300 μl of the frozen plasma using a 3D-Gene® RNA extraction reagent (TORAY Industries, Inc., Tokyo, Japan) and the half of the extracted product was used. After being labeled with an Exiqon miRCURY LNA™ microRNA Array Power Labeling kit (Exiqon Inc., Woburn, MA, USA), miRNAs were analyzed with a 3D-Gene® Human miRNA oligo chip (Ver. 17.0) (TORAY Industries, Inc., Tokyo, Japan) which carries approximately 1800 probes to detect human miRNAs using fluorescent signals. From the outputted signal data, we subtracted the background noise, and calibrated using the global normalization method with the median value as 25.
Comparisons of all normalized miRNA data calculated as a binary logarithm underwent a paired t test, Wilcoxon rank sum test, fold change ratio and variance using JMP® Pro 11.0.0. (SAS Institute Inc., Cary, North Carolina, USA). MiRNA showing the most significant changes either up-regulating or down-regulating in sALS patients, were chosen as the biomarker candidates for validation analysis.

Takahashi I., Hama Y., Matsushima M., Hirotani M., Kano T., Hohzen H., Yabe I., Utsumi J, & Sasaki H. (2015). Identification of plasma microRNAs as a biomarker of sporadic Amyotrophic Lateral Sclerosis. Molecular Brain, 8, 67.

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Profiling Endothelial Cell miRNAs

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After concentration adjustment and placement into 24-well plates at 1 × 10⁴ cells/well, PMVECs were subjected to the grouped treatments mentioned above and a 24 h routine culture in the 5% CO₂ incubator at 37°C after mixing gently. Firstly, total RNA in PMVECs was sequestered by the use of TRIzol and further was highly purified by virtue of RNeasy kits. Then, a UV spectrophotometer was wielded to measure the RNA level. Secondly, miRCURY™ LNA microRNA Array Power Labeling Kit (Exiqon) was utilized to label the total RNA in cell samples. The results after hybridization were scanned using the GeneChip Scanner 3000 system, and Command Console software v4.0 (Affymetrix) was introduced for reading raw data. Thirdly, after normalization, the differentially expressed miRNAs in the three groups were screened by fold change >2.0 and examined by t-test. The differentially expressed miRNAs screened in each group were intuitively displayed by cluster maps [18 (link)].

Liu Z, & Yang Y. (2022). Ginkgolide A Participates in LPS-Induced PMVEC Injury by Regulating miR-224 and Inhibiting p21 in a Targeted Manner. Contrast Media & Molecular Imaging, 2022, 6384334.

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Profiling miRNA Expressions in Liver Cancer

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miRCURY LNA™ microRNA chips (version 8.0, Exiqon, Vedbaek, Denmark) were used to profile the differences for miRNA expressions among SLHCC, SHCC and NHCC. The array contained a total of 840 specific probes in triplicates. It was performed according to the protocol of miRCURY LNA microRNA Array Power Labeling kit (Exiqon) [34 (link)]. The image analysis was conducted in Genepix Pro 6.0 (Axon Instruments) as described before [35 (link)] (details were described in the Supplementary Materials and Methods).

Yang H., Zheng W., Shuai X., Chang R.M., Yu L., Fang F, & Yang L.Y. (2015). MicroRNA-424 inhibits Akt3/E2F3 axis and tumor growth in hepatocellular carcinoma. Oncotarget, 6(29), 27736-27750.

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Profiling microRNA Expression in Type 1 Diabetes

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Two-color hybridization was performed with miRCURY LNA microRNA arrays (7th generation [miRBase v18]; Exiqon, Vedbaek, Denmark), co-hybridizing total RNA from each type 1 diabetic mouse (n = 6) with total RNAs from a pool of 6 non-diabetic animals.
1.25 ug of total RNA was labeled with either Hy3 (non-diabetic pool) or Hy5 (diabetic) using the miRCURY LNA microRNA Array Power labeling kit (Exiqon), according to the manufacturer's protocol. Additionally, two dye-swaps were performed, labeling the diabetic sample with Hy3 and the control pool with Hy5.
Hybridization at 53 C for 16 h and washing steps were carried out on the HS 400 PRO hybridization station (Tecan, Männedorf, Switzerland). MicroRNA arrays were scanned using the GenePix 4100A microarray scanner and the GenePix Pro GenePix Pro 6.1.0.4 software (Molecular Devices, San Jose, CA, USA).

Baldini E., Testa E., Voellenkle C., De Domenico E., Cianfarani F., Martelli F., Ulisse S, & Odorisio T. (2020). Dysregulation of microRNA expression in diabetic skin. Journal of dermatological science, 98(3).

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Comprehensive mRNA and miRNA Expression Profiling

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mRNAs were labeled with Cy5 mono-reactive dye (GE Healthcare, Little Chalfont, UK) and purified according to the manufacturer's protocol (3DGENE mRNA CyDye label v2, Toray Industries, Tokyo, Japan). The concentration of labeled RNAs was determined by a NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific) before hybridization onto microarray chips (Human Oligo Chip 25k ver. 2.10., Toray Industries). Hybridization and subsequent washing were performed according to the manufacturer's protocol (3DGENE mRNA hybridization v2, Toray Industries). For miRNA analysis, miRNA was labeled, hybridized, and washed according to the manufacturer's protocol (3DGENE miRNA label Hybridization 4plex v1, Toray Industries) using the miRCURY LNA microRNA Array Power Labeling kit (Exiqon, Vedbaek, Denmark) and Human miRNA oligo chip (Human miRNA Chip ver. 16.1.0.0, Toray Industries). The intensity of labeled mRNAs or miRNAs was analyzed with the 3D-Gene Scanner 3000 system with auto gain, auto focus, and auto analysis settings.

Ito M., Sun S., Fukuhara T., Suzuki R., Tamai M., Yamauchi T., Nakashima K., Tagawa Y.I., Okazaki S., Matsuura Y., Wakita T, & Suzuki T. (2017). Development of hepatoma-derived, bidirectional oval-like cells as a model to study host interactions with hepatitis C virus during differentiation. Oncotarget, 8(33), 53899-53915.

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