MiRNA array analysis was performed as described in our previous study [18 (link)]. Total RNA was extracted from tissue samples using a miRNAeasy Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. MiRNA array was performed using a Human miRNA Oligo Chip (v. 21.0; Toray Industries, Tokyo, Japan). Total miRNA samples (200 ng) were labeled using a miRCURYHy3/Hy5 Power Labeling Kit, slides were scanned in a 3D-Gene Scanner 3000 (Toray Industries), and the fluorescence data were analyzed using 3D-Gene Extraction software (v. 1.2; Toray Industries). Quantile normalization was performed on raw data exceeding the background level, and differentially expressed miRNAs were identified by the Mann–Whitney U test. Hierarchical clustering was performed using the farthest end method, with the absolute non-central Pearson correlation coefficient as the metric. A heat map was created based on the relative expression intensity of each miRNA, using log2 of the fold change.
Mircuryhy3 hy5 power labeling kit
Manufactured by Toray
Sourced in Japan
The MiRCURYHy3/Hy5 Power Labeling Kit is a laboratory equipment product designed for the labeling of miRNA samples. It provides the necessary reagents and materials for the preparation of fluorescently labeled miRNA samples for use in various downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
3d gene scanner 3000, by Toray (4 mentions)
3d gene extraction software, by Toray (2 mentions)
Mirneasy mini kit, by Qiagen (2 mentions)
3d gene extraction version 1, by Toray (2 mentions)
Human mirna oligo chip, by Toray (1 mentions)
Mirnaeasy mini kit, by Qiagen (1 mentions)
Genespring gx, by Agilent Technologies (1 mentions)
Mirneasy serum plasma kit, by Qiagen (1 mentions)
Genespring gx v10, by Agilent Technologies (1 mentions)
Mirneasy kit, by Qiagen (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Rna 6000 nano kit, by Agilent Technologies (1 mentions)
4 protocols using mircuryhy3 hy5 power labeling kit
1
miRNA Array Analysis Protocol
2
Exosomal miRNA profiling after ED-71 treatment
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We examined the effect of ED-71 on the miRNA profile of A431-derived exosomes. Total RNA was extracted from purified exosomes using miRNeasy Serum/Plasma Kit (QIAGEN) according to manufacturer's instructions. Samples were labeled using a miRCURYHy3/Hy5 Power Labeling Kit and were hybridized to a human miRNA Oligo chip (Toray Industries, Tokyo, Japan). The chips were scanned with a 3D-Gene Scanner 3000 (Toray Industries). The results were analyzed using 3D-Gene extraction software (Toray Industries). Differences in miRNA expression between ED-71-treated and control cells were assessed using GeneSpring GX (Agilent Technologies, Santa Clara, CA). Quantile normalization was performed on the raw data that were above the background level.
3
Quantification of Liver Cancer miRNA
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Total miRNA was extracted from liver cancer cell lines using the Qiagen miRNeasy kit (Qiagen K.K.) according to the instructions provided by the manufacturer. Each serum sample was processed for total RNA extraction with the miRNeasy Mini Kit (Qiagen) according to the manufacturer's instructions. The RNA sample from both sets typically showed A260/280 ratios between 1.9 and 2.1 measured with an Agilent 2100 Bioanalyzer (Agilent Technologies).
After RNA quantification with an RNA 6000 Nano kit (Agilent Technologies), the samples were labeled using a miRCURY Hy3/Hy5 Power labeling kit and hybridized on a human miRNA Oligo chip10, version 14.0 (Toray Industries). Scanning was conducted with the 3D-Gene Scanner 3000 (Toray Industries). The 3D-Gene extraction version 1.2 software (Toray Industries) was used to read the raw intensity of the image. The raw data were analyzed with GeneSpringGX v 10.0 (Agilent Technologies) to determine the change in miRNA expression. The samples were frozen at −80°C within 4 h of collection and thawed just before analysis.
After RNA quantification with an RNA 6000 Nano kit (Agilent Technologies), the samples were labeled using a miRCURY Hy3/Hy5 Power labeling kit and hybridized on a human miRNA Oligo chip10, version 14.0 (Toray Industries). Scanning was conducted with the 3D-Gene Scanner 3000 (Toray Industries). The 3D-Gene extraction version 1.2 software (Toray Industries) was used to read the raw intensity of the image. The raw data were analyzed with GeneSpringGX v 10.0 (Agilent Technologies) to determine the change in miRNA expression. The samples were frozen at −80°C within 4 h of collection and thawed just before analysis.
4
MiRNA Expression Analysis Protocol
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MiRNA array analysis was performed as described in our previous study.[ 21 ] Briefly, total RNA was isolated using a miRNeasy Mini Kit (Qiagen, Hilden, Germany) from PK-1 cells according to the manufacturer's instructions. MiRNA expression analysis was performed using the miRCURY Hy3/Hy5 Power Labeling Kit and human miRNA Oligo chip (v. 21.0; Toray Industries, Tokyo, Japan). The arrays were scanned in a 3D-Gene Scanner 3000 (Toray Industries), and these fluorescence images were analyzed using the 3D-Gene extraction version 1.2 software (Toray Industries).
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