Mircury lna mirna array

Manufactured by Qiagen

Sourced in Denmark

The MiRCURY LNA miRNA Array is a high-performance microarray platform designed for the comprehensive analysis of microRNA (miRNA) expression. The array utilizes Locked Nucleic Acid (LNA) technology to provide sensitive and specific detection of miRNA molecules. The core function of this product is to facilitate the global profiling and quantification of miRNA expression levels in biological samples.

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Lab products found in correlation

Genepix pro 6, by Molecular Devices (3 mentions) Axon genepix 4000b microarray scanner, by Molecular Devices (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Wash buffer kit, by Qiagen (1 mentions) Mircury array power labeling kit, by Qiagen (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Nd 1000, by Thermo Fisher Scientific (1 mentions) Axon genepix 4000b, by Molecular Devices (1 mentions) Mirneasy mini kit, by Qiagen (1 mentions) Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Lightcycler 480 program, by Roche (1 mentions) Taqman microrna assay kit, by Thermo Fisher Scientific (1 mentions) Nanodrop nd 2000 spectrophotometer, by Thermo Fisher Scientific (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) Qiazol reagent, by Qiagen (1 mentions) Genespring gx, by Agilent Technologies (1 mentions) Agilent feature extraction software v10.7, by Agilent Technologies (1 mentions) Agilent g2565ba microarray scanner system, by Agilent Technologies (1 mentions) Imagene 9, by BioDiscovery (1 mentions) Agilent 2100 bioanalyzer profile, by Agilent Technologies (1 mentions)

8 protocols using mircury lna mirna array

Microarray Analysis of LEC Transcriptome

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The preparation of RNA samples and microarray analysis were performed commercially
by RiboBio Co. Ltd (Guangzhou, China). In brief, total RNA of LECs treatment with
or without TGFβ2 was isolated using TRIzol (Invitrogen, Carlsbad,
CA, USA) and miRNeasy mini kit (QIAGEN, Hilden, Germany) according to the
manufacturer’s instructions, which efficiently recovers all RNA species,
including miRNAs. RNA quality and quantity were measured by nanodrop
spectrophotometer (ND-1000, Nanodrop Technologies, Wilmington, DE, USA), and RNA
integrity was determined by gel electrophoresis. The isolated miRNAs were then
labeled with Hy3/Hy5 using the miRCURY Array Power Labeling kit (Exiqon,
Vedbaek, Denmark) and hybridized on a miRCURY LNA miRNA Array (v.18.0, Exiqon)
according to array manual. Following hybridization, the slides were achieved,
washed several times using wash buffer kit (Exiqon) and finally dried by
centrifugation for 5 min at 400 r.p.m. Then the slides were scanned using
the Axon GenePix 4000B microarray scanner (Axon Instruments, Foster City, CA,
USA). The scanned images were then imported into GenePix Pro 6.0 software (Axon
Instruments) for grid alignment and data extraction. Bioinformatics analysis and
visualization of microarray data were performed with MEV software (v4.6, TIGR).
The microarray assays were repeated three times each group.

Chen X., Xiao W., Chen W., Liu X., Wu M., Bo Q., Luo Y., Ye S., Cao Y, & Liu Y. (2017). MicroRNA-26a and -26b inhibit lens fibrosis and cataract by negatively regulating Jagged-1/Notch signaling pathway. Cell Death and Differentiation, 24(8), 1431-1442.

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Selection of Inflammation-related miRNAs

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MiRNA were selected based on literature search. We used “inflammation”, “chronic stress”, and “resilience” as search terms and cross-compared results for miR103a-3p, miR107, miR130a-3p, and miR125a-5p in public data bases (miRbase [32 ], NCBI [33 ]). We then compared our results with those of a previous miRCURY LNA miRNA array taken by the profiling service of Exiqon (Exiqon Services, Vedbaek, Denmark) [9 (link)] and decided on four miRNA that had survived Benjamini-Hochberg correction (p < 0.05) and showed ≥ 60fold log-fold change.

Braun A., Evdokimov D., Frank J., Sommer C, & Üçeyler N. (2020). MiR103a-3p and miR107 are related to adaptive coping in a cluster of fibromyalgia patients. PLoS ONE, 15(9), e0239286.

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Transgenic Mice miRNA Profiling

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Equal amounts of RNA samples from the transgenic and WT mice (n = 6) were subjected to miRNA profiling using a miRCURY LNA miRNA array (v11.0; Exiqon, Vedbaek, Denmark).

Lyu Q.L., Jiang B.M., Zhou B., Sun L., Tong Z.Y., Li Y.B., Tang Y.T., Sun H., Liu M.D, & Xiao X.Z. (2018). MicroRNA Profiling of Transgenic Mice with Myocardial Overexpression of Nucleolin. Chinese Medical Journal, 131(3), 339-346.

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miRNA Expression Profiling by Microarray

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The 5 groups of samples were separately labeled using the miRCURY LNA miRNA Hy3/Hy5 Power Labeling kit (208030-A; Exiqon, Woburn, MA, USA) and hybridized on the miRCURY LNA miRNA array (version 14.0; 5th Gen Human; Exiqon). After the washing steps, the slides were scanned using the Axon GenePix 4000B Microarray scanner (Molecular Devices, Sunnyvale, CA, USA). Scanned images were then imported into GenePix Pro 6.0 software (Axon; Molecular Devices) for grid alignment and data extraction. Replicated miRNAs were averaged and miRNAs with intensities >50 in all samples were selected for calculating the normalization factor. Expressed data were normalized using the median. Following normalization, differentially expressed miRNAs were identified through fold change filtering.

LI S.Q., LI F., XIAO Y., WANG C.M., TUO L., HU J., YANG X.B., WANG J.S., SHI W.H., LI X, & CAO X.F. (2014). Comparison of long non-coding RNAs, microRNAs and messenger RNAs involved in initiation and progression of esophageal squamous cell carcinoma. Molecular Medicine Reports, 10(2), 652-662.

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Profiling Cardiac miRNA Expression

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Total RNA was harvested from heart tissue and cardiomyocytes using Qiazol reagent (Qiagen, Valencia, CA, USA), according to the manufacturer’s instructions. The concentration of each sample was measured using a Nanodrop ND-2000 spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, DE, USA). For miRNA microarray analysis, miRNA expression profiling was conducted by miRNA microarray analysis using the miRCURY LNA™ miRNA Array (Exiqon, Vedbaek, Denmark) containing 700 mature rat miRNAs. The accuracy of the microarray data was validated using real-time PCR with TaqMan probes. For real-time PCR analysis, total RNA was reverse transcribed with stem-loop primers and the TaqMan® MicroRNA Reverse Transcription kit (Applied BioSystems, Foster City, CA, USA), according to the manufacturer’s instructions [28 (link)]. Real-time PCR was performed in duplicate using the TaqMan® MicroRNA assay kit and TaqMan Universal PCR MasterMix (Applied Biosystems) for miRNA-23a and miRNA-92a, according to the manufacturer’s instructions. Real-time PCR was performed using the LightCycler® 480 program (Roche) for 40 cycles, (10 s each, at 95°C, 60°C, 72°C). Relative miRNA expression levels were normalized using the RNU6B (U6) small non-coding RNA as an endogenous control. The information of TaqMan® MicroRNA assay kit is shown in S1 Table.

Song Y.S., Joo H.W., Park I.H., Shen G.Y., Lee Y., Shin J.H., Kim H, & Kim K.S. (2017). Bone marrow mesenchymal stem cell-derived vascular endothelial growth factor attenuates cardiac apoptosis via regulation of cardiac miRNA-23a and miRNA-92a in a rat model of myocardial infarction. PLoS ONE, 12(6), e0179972.

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Fecal miRNA Microarray Profiling

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Kang Chen (Shanghai, China) performed the miRNA microarray screening after RNA isolation from the specimens. Total miRNAs from fecal specimens were labeled with miRCURY™ Hy3™/Hy5™ fluorescent dye using the miRCURY™ Power Labelling Kit (Exiqon, Vedbaek, Denmark) following the manufacturer’s guidelines, and then hybridized to the miRCURY™ LNA miRNA Array (version 18.0, Exiqon, Vedbaek, Denmark). The Axon GenePix 4000B microarray scanner (Molecular Devices, USA) was used to scan the slides. The Axon GenePix 4000B microarray scanner (Molecular Devices, USA) was used to scan the slides. The data were extracted with Agilent Feature Extraction v10.7 software and analyzed with GeneSpring GX (version 13.0, Agilent Technologies, California, USA). After normalization, the absolute value of the fold change of gene expression was more than seven as a significant difference, and this method was used to screen DEMs.

Zhou R., Qiu P., Wang H., Yang H., Yang X., Ye M., Wang F, & Zhao Q. (2021). Identification of microRNA-16-5p and microRNA-21-5p in feces as potential noninvasive biomarkers for inflammatory bowel disease. Aging (Albany NY), 13(3), 4634-4646.

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Profiling miRNA Expression Using Microarray

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All experiments were conducted at Exiqon Services. The quality of the total RNA was verified by an Agilent 2100 Bioanalyzer profile. Total RNA (250 ng) from sample and reference was labeled with Hy3 and Hy5 fluorescent label, respectively, using the miRCURY LNA miRNA Hi-Power Labelling Kit, Hy3/Hy5 (Exiqon, Denmark), following the procedure described by the manufacturer. The Hy3- labeled samples and a Hy5- labeled reference RNA sample were mixed pairwise and hybridized to the miRCURY LNA miRNA Array (sixth generation: hsa, mmu, and rno) (Exiqon, Denmark), which contains capture probes targeting all miRNAs for human, mouse, or rat registered in the miRBASE 16.0. The hybridization was performed according to the miRCURY LNA miRNA Array instruction manual using a Tecan HS 4800 hybridization station (Tecan, Austria). After hybridization, the microarray slides were scanned and stored in an ozone-free environment (ozone level below 2.0 parts per billion) to prevent potential bleaching of the fluorescent dyes. The miRCURY LNA array microarray slides were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies Inc.), and the image analysis was carried out using the ImaGene 9.0 software (BioDiscovery Inc.). The quantified signals were background-corrected (Normexp with an offset value of 10) and normalized using the Quantile normalization algorithm.

Schmolka N., Papotto P.H., Romero P.V., Amado T., Enguita F.J., Amorim A., Rodrigues A.F., Gordon K.E., Coroadinha A.S., Boldin M., Serre K., Buck A.H., Gomes A.Q, & Silva-Santos B. (2018). MicroRNA-146a controls functional plasticity in γδ T cells by targeting NOD1. Science immunology, 3(23), eaao1392.

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Profiling miRNA Expression in Hypoxic Cells

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Total RNA from HCT116 and SW480 cells cultured under hypoxic or normoxic conditions was extracted using the TRIzol reagent (Thermo Fisher Scientific), and the miRNA part was further purified using a mirVana miRNA extraction kit (Ambion, Austin, TX, USA). The isolated miRNAs were labeled with Hy3 using a miRCURY microarray labeling kit (Exiqon) and then hybridized using a miRCURY LNA miRNA Array (v.8.0, Exiqon). The microarray data were collected using a GenePix 4000B scanner (Axon Instruments, Molecular Devices, San Jose, CA, USA) and analyzed using a GenePix Pro 6.0 software (Axon Instruments).

Tang Y., Weng X., Liu C., Li X, & Chen C. (2021). Hypoxia Enhances Activity and Malignant Behaviors of Colorectal Cancer Cells through the STAT3/MicroRNA-19a/PTEN/PI3K/AKT Axis. Analytical Cellular Pathology (Amsterdam), 2021, 4132488.

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