The same amounts of RNA from different animals in each group were pooled (n = 5) as one microarray sample and used for poly A tailing and ligation with the FlashTag Biotin HSR RNA Labeling Kit (Affymetrix, Santa Clara, CA, USA). The labeled samples were mixed with a hybridization cocktail (2×Hybridization Mix, 27.5% formamide, DMSO, 20× Eukaryotic Hybridization Controls, Control Oligonucleotide B2 from GeneChip Eukaryotic Hyb Control Kit, Affymetrix) and hybridized onto an miRNA 2.0 Array (Affymetrix) at 48 °C for 16 h. Thereafter, washing and staining were performed using an Affymetrix GeneChip Fluidics Station 450. Scans were performed using an Affymetrix GeneChip Scanner 3000. Data analysis was performed using the Affymetrix miRNA Array QC tool. Each experiment was performed following the manufacturer’s instructions.
Mirna 2.0 array
Manufactured by Thermo Fisher Scientific
Sourced in United States
The MiRNA 2.0 array is a microarray platform designed for the detection and analysis of microRNA (miRNA) expression levels. It provides comprehensive coverage of human, mouse, and rat miRNA sequences. The array enables the simultaneous measurement of thousands of miRNA species in a single experiment, allowing for the profiling of miRNA expression patterns across diverse biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Genechip fluidics station 450, by Thermo Fisher Scientific (2 mentions)
Genechip scanner 3000, by Thermo Fisher Scientific (2 mentions)
Flashtag biotin hsr rna labeling kit, by Thermo Fisher Scientific (1 mentions)
Eukaryotic hybridization controls, by Thermo Fisher Scientific (1 mentions)
Mirvana isolation kit, by Thermo Fisher Scientific (1 mentions)
Rneasy total rna isolation kit, by Qiagen (1 mentions)
3 ivt express kit, by Thermo Fisher Scientific (1 mentions)
Mirvana mirna isolation kit, by Thermo Fisher Scientific (1 mentions)
Genechip hybridization oven 640, by Thermo Fisher Scientific (1 mentions)
Mouse gene 1.0 st array, by Thermo Fisher Scientific (1 mentions)
Dynabead, by Thermo Fisher Scientific (1 mentions)
7 protocols using mirna 2.0 array
1
miRNA Profiling of Animal Samples
2
mRNA and miRNA Microarray Analysis of 3T3-L1 Cells
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For mRNA microarray, total RNA of 3T3-L1 cells was isolated with an RNeasy Total RNA Isolation kit (QIAGEN). The overall gene-expression profiles were detected by Affymetrix Mouse 430 2.0 according to Affymetrix GeneChip expression-assay protocols. For miRNA microarray, total RNA of 3T3-L1 cells was isolated by using miRVana Isolation Kit according to the manufacturer’s instructions (Ambion). The miRNA expression profiles were detected by Affymetrix miRNA 2.0 Array. Three independent experiments and two independent experiments were done for mRNA and miRNA (We originally had three biological repeats on miRNA array, but one of the sample had problems, so we only selected two biological repeats for further analysis), respectively.
3
Profiling miRNA Expression in OC3 Cells
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The miRNA expression levels between OC3 and OC3-IV2 cells were analyzed using Affymetrix miRNA 2.0 array (Affymetrix, Santa Clara, CA, USA). The experiment was performed at Microarray Core Laboratory, National Health Research Institute, Taiwan.
4
Profiling miRNA Expression in Breast Cancer Metastasis
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The miRNA expression pattern between the MDA-MB-231 parental cells and three in vivo-selected lung metastatic IV2 sublines were analyzed using Affymetrix miRNA 2.0 array (Affymetrix, Santa Clara, CA, USA). The miRNA array experiment was carried out at Microarray Core Laboratory in National Health Research Institute (Miaoli, Taiwan).
5
miRNA Expression Profiling Using Affymetrix Arrays
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Total RNA from cultured cells, with efficient recovery of small RNA, were isolated using a mirVana™ miRNA Isolation kit (Ambion, Austin, TX, USA). cDNA for each sample was synthesized by using the 3′ IVT Express kit (Affymetrix, Santa Clara, CA, USA) according to the supplier's protocol. The purified cDNA was fragmented by incubation in fragmentation buffer (provided in the 3′IVT Express kit) at 95°C for 35 min and chilled on ice. The fragmented labeled cDNA was subjected to an miRNA 2.0 array (Affymetrix) and hybridized in a Genechip hybridization oven 640 (Affymetrix) at 45°C for 18 h. After washing and staining in a GeneChip Fluidics Station 450 (Affymetrix), the arrays were scanned by using GeneChip Scanner 3000 (Affymetrix). The gene expression levels of samples were normalized and compared by using Partek Genomics Suite 6.5 (Partek Inc., St. Louis, MO, USA). Average-linkage hierarchical clustering of the data was applied by using the Cluster function and the results were displayed by using TreeView (http://genome-www.stanford.edu/clustering/ ).
6
Amygdalae miRNA Profile Analysis
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Pools of three amygdalae taken from three mice from the same treatment group (either social defeat, n = 18, or control, n = 12) were immunoprecipitated using magnetic protein G beads (Dynabeads, Invitrogen/Life Technologies) and Ago2 monoclonal antibody (WAKO Chemicals).
RNA from the Ago2 IP samples was isolated and analyzed on an Affymetrix miRNA 2.0 array (enriched RNA protocol) and an Affymetrix Mouse Gene 1.0 ST array.
RNA from the Ago2 IP samples was isolated and analyzed on an Affymetrix miRNA 2.0 array (enriched RNA protocol) and an Affymetrix Mouse Gene 1.0 ST array.
7
COPD Patient Lung Tissue Analysis
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Lung tissue samples from COPD patients were obtained through our tissue bank in collaboration with the Division of Thoracic Surgery at the University Hospital of Zurich. In the present study patients with severe COPD that fulfilled the Global Initiative for COPD (GOLD) criteria were included. Written informed consent was obtained from all patients. The study design was approved by Zurich's ethical review committee (KEK ZH-Nr 2014-0011).
Validation of human gene expression was performed in patient samples derived from the GLUCOLD study cohort measured by Affymetrix Human Gene ST array and Affymetrix miRNA 2.0 array (GSE36221) [12] [13] (link)[14] (link).
Validation of human gene expression was performed in patient samples derived from the GLUCOLD study cohort measured by Affymetrix Human Gene ST array and Affymetrix miRNA 2.0 array (GSE36221) [12] [13] (link)[14] (link).
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