Twenty RNA samples [6 normal thyroid samples, 6 papillary thyroid cancer (PTC), and 8 anaplastic thyroid cancer (ATC)] were labeled using miRCURY LNA microRNA Hi-Power Labeling Kit (Exiqon). The labeled samples were hybridized to the miRCURY LNA microRNA Arrays. The dataset from Exiqon was loaded into Partek Genomics Suite (v6.6; Partek Inc.). To perform quality control and statistical analysis, one-way ANOVA was applied in the analysis with two linear contrasts. One contrast was to compare ATC to normal and another contrast was to compare ATC with PTC. The fold change was set at > ±1.5, false discovery rate (adjusted P value) <0.05. Differential expression (DE) of miRNAs between ATC and normal was 64 and DE miRNAs between ATC and PTC was 93. The intersection of DE miRNAs was 31. The 31 differentially expressed miRNAs are shown in Fig. 1A by Hierarchical Clustering.
Mircury lna microrna hi power labeling kit
Manufactured by Qiagen
Sourced in Denmark
The MiRCURY LNA microRNA Hi-Power Labeling Kit is a laboratory equipment product designed for the efficient labeling of microRNA samples. The kit provides the necessary reagents and protocols to facilitate the labeling process, allowing researchers to prepare their microRNA samples for downstream analysis.
Automatically generated - may contain errors
Lab products found in correlation
Mircury extraction kits, by Qiagen (2 mentions)
Mircury lna microrna array, by Qiagen (2 mentions)
Agilent g2565ba microarray scanner system, by Agilent Technologies (2 mentions)
Hs4800 hybridization station, by Tecan (2 mentions)
Mircury lna microrna array 7th gen, by Qiagen (2 mentions)
Imagene 9, by BioDiscovery (1 mentions)
G2505c dna microarray scanner, by Agilent Technologies (1 mentions)
Surehyb chamber, by Agilent Technologies (1 mentions)
Genepix 4000b microarray scanner, by Molecular Devices (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Feature extraction 10, by Agilent Technologies (1 mentions)
Agilent 2100 bioanalyzer profile, by Agilent Technologies (1 mentions)
Mircury rna isolation kit tissue, by Qiagen (1 mentions)
Rnalater, by Thermo Fisher Scientific (1 mentions)
Imagene 9 mircury lna microrna array analysis software, by Qiagen (1 mentions)
10 protocols using mircury lna microrna hi power labeling kit
1
Differential miRNA Expression in Thyroid Cancer
2
ApoE Isoform-Specific Effects on Neuronal miRNAs
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Embryonic 17 days old ApoE−/− cortical neurons were cultured for 7 days in vitro in the presence of conditioned media derived from ApoE3 and ApoE4 primary astrocyte cultures as described [26 (link), 43 (link)] (N = 3/group). miRNAs were extracted using miRCURY extraction kits (Exiqon Inc.) and then labeled using miRCURY LNA microRNA Hi-Power Labeling kit, Hy3/Hy5, and hybridized on the miRCURY LNA microRNA Array. The quality assessment using control spike-in oligo nucleotides produced signals in expected range indicated successful labeling. Following normalization of quantified signals after background correction using global Lowess regression algorithm, unsupervised and supervised data analysis were performed. The microRNA profiling identified a subset of microRNAs that are differentially expressed in the ApoE3 versus ApoE4-treated neurons.
3
miRNA Microarray Screening of Endometriosis
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For the purpose of miRNA microarray screening we chose 10 samples of patients with advanced endometriosis and 11 controls. Microarray analysis was conducted as single-channel Hy3 experiments on Exiqon's miRCURY LNA (locked-nucleic acid) microRNA Array 7th generation—hsa, mmu and rno. Exiqon arrays contain 3100 capture probes, complementary to most human, mouse, rat, and their related viral sequences from the v.19.0 release of miRBase. The arrays also contain 25 proprietary human miRPlus sequences not yet in miRBase. 500 ng RNA sample was labelled with a Hy3 fluorophore (Exiqon, Denmark). Labelling reactions were performed using Exiqon's miRCURY LNA microRNA Hi-Power Labeling Kit with the use of synthetic spike controls, Spike-in microRNA Kit v2 (Exiqon, Denmark), according to the manufacturer's protocol. Hybridization of labeled RNA to the array was performed in SureHyb chambers (Agilent Technologies, USA) for 16 hours at 56°C. Slides were washed according to manufacturer's instructions and scanned at 10 μm resolution using an Agilent G2505C DNA Microarray Scanner. Raw data were generated using Imagene 9.0 software (BioDiscovery, Inc., USA), using an FE protocol available on demand from Exiqon.
4
ApoE Isoform Effects on Cortical Neuron miRNA
5
Microarray-Based miRNA Profiling Protocol
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Microarrays were generated by spotting Nexterion HiSens E Sildes (Schott, ordered by Peqlab, Prod. no. 39-1125813, Erlangen, Germany) with the miRCURY LNA microRNA array ready-to-spot probe set, 6th gen, human, mouse & rat (Exiqon, prod. no. 208410, Vedbaek, Denmark). This probe set consists of 2383 unique capture probes, covering all mature miRNAs from human, mouse and rat annotated in miRBase version 16.0. For two-colour hybridisations, 1 μg total RNA of each sample was used for Hy3 and Hy5 labelling using the miRCURY LNA microRNA Hi-Power Labeling Kit (Exiqon, Prod. no. 208035) according to the manufacturer's instructions. Hybridisations were performed on a TECAN HS400 Hybridisation Station (Tecan, Männedorf, Switzerland). All hybridisations were repeated with reversed dye assignment (dye-swap). Hybridised slides were scanned with a GenePix 4000B microarray scanner (Molecular Devices, Sunnyvale, CA, USA) at 10 μm resolution and the resultant TIFF images were analysed with GenePix Pro 4.1 software. Raw data were subsequently processed using ArrayNorm (Pieler et al, 2004 (link)) for normalisation and Genesis software for evaluation (Sturn et al, 2002 (link)). All experimental parameters, raw and processed data files were submitted to the public repository Gene Expression Omnibus (GEO accession GSE53095).
6
Profiling microRNA Expression in Ocular and Lymphatic Tissues
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Total RNA was isolated from iris, ciliary bodies, and popliteal lymph nodes with Trizol reagent (Life Technologies, Gaithersburg, MD, USA). The samples were labeled using the miRCURY LNA microRNA Hi-Power Labeling Kit, Hy3/Hy5, and hybridized on the miRCURY LNA microRNA Array (7th Gen, Exiqon, Vedbæk, Denmark) in accordance with the manufacturer's instructions. Image analysis was then performed to quantify the signals on the array. The mean ± standard deviation was calculated for each group, and miRNA signals were all transformed to logarithm base 2 for further statistical analysis.
7
miRNA Profiling and Analysis
8
Microrna Profiling of Fracture Repair
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For miRNA microarray analysis, tissues were harvested on post-fracture day 14 and stored in RNAlater (Ambion, Austin, TX, USA). Total RNA, including miRNA, was extracted from the tissue specimens of five different animals in each group using a miRCURY RNA Isolation Kit-Tissue (Exiqon, Vedbaek, Denmark). The quality of the total RNA was verified using an Agilent 2100 Bioanalyzer profile (Agilent Technologies, Santa Clara, CA, USA). One microgram of total RNA from sample and reference RNAs was labeled with Hy3 and Hy5 fluorescent labels, respectively, using a miRCURY LNA microRNA Hi-Power Labeling Kit (Exiqon). The Hy3-labeled samples and a Hy5-labeled reference RNA sample were mixed pair-wise and hybridized to a miRCURY LNA Array, version 6th Generation (Exiqon), which contains capture probes targeting all human, mouse, and rat miRNAs registered in miRBASE version 16.0. Labeling, hybridization, washing, and scanning were performed following the instructions of the manufacturers. Data analysis was carried out using Feature Extraction 10.7.3.1 (Agilent Technologies).
9
Exiqon microRNA microarray profiling
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The microarray experiment using the Exiqon platform was performed by Exiqon (Denmark). Briefly, 0.75 μg of total RNA from samples were labeled using the miRCURY LNA™ microRNA Hi-Power Labeling Kit, Hy3™/Hy5™ and hybridized on the miRCURY LNA™ microRNA Array 7th Gen (Exiqon, Denmark) using a Tecan HS4800™ hybridization station (Tecan, Austria). After hybridization the microarray slides were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., USA) and the image analysis was carried out using the ImaGene® 9 (miRCURY LNA™ microRNA Array Analysis Software, Exiqon, Denmark).
10
Differential miRNA Expression in TNF-α-Treated NPCs
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NPCs from three randomly selected samples (5 × 106 per sample) with and without treatment of TNF‐α were harvested as the test and reference sample, respectively. Total RNA was labelled using the miRCURY LNA microRNA Hi‐Power Labeling Kit, Hy3/Hy5, and hybridized on the miRCURY LNA microRNA Array (Exiqon, Vedbæk, Denmark). Three independent hybridizations for each sample were performed on chip. After hybridization, the microarray slides were scanned using GenePix 4000B scanner (Axon, Foster City, CA, USA) and the data were analysed using GenePix Pro V6.0 software (Axon). Expression data were normalized using the locally weighted scatter plot smoothing (LOWESS) regression algorithm. miRNAs with twofold difference and statistical significance between groups were considered as differentially expressed. Hierarchical cluster and heat map analyses were performed using the MultiExperiment Viewer v4.8 program of TM4 Microarray Software Suite. The DIANA‐mirPath v3.0 database was used to identify target genes and pathways potentially altered by the down‐regulated miRNAs in TNF‐α‐treated NPCs. The software created a clustering of the selected miRNAs based on their influence on molecular pathways 30 .
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