Total RNA was extracted from three eutopic and three ectopic endometrium tissues by miRNeasy isolation kit (Qiagen, Inc.) according to the manufacturer's protocol. Total RNA (200 ng) was labeled with fluorescence dye Hy3 or Hy5 using the miRCURY Hy3/Hy5 Power Labeling kit (Exiqon). After hybridization on the miRCURY™ LNA Array (v.18.0, Exiqon), data obtained from Axon GenePix 4000B microarray scanner (Axon Instruments; Molecular Devices, LLC) were imported into the GenePix Pro 6.0 program (Axon Instruments; Molecular Devices, LLC) for analysis (15 (link)). The bioinformatics analysis was performed by RiboBio Co., Ltd. Finally, the heatmap of miRNAs with the most marked differences was created using hierarchical clustering in GeneSpring GX software, version 7.3 (Agilent Technologies, Inc.).
Protocol detail
Differential miRNA Expression in Endometriosis
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Corresponding Organization :
Other organizations : Lanzhou University, Lanzhou University Second Hospital, Northwest Women's and Children's Hospital, Dongguan People’s Hospital
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Variable analysis
independent variables
- Tissue type (eutopic endometrium, ectopic endometrium)
- MiRNA expression levels
- Total RNA amount (200 ng) for labeling
- Fluorescence dyes used for labeling (Hy3 or Hy5)
- MiRCURY LNA Array (v.18.0) used for hybridization
- Axon GenePix 4000B microarray scanner used for data acquisition
- GenePix Pro 6.0 program used for data analysis
- GeneSpring GX software (version 7.3) used for hierarchical clustering and heatmap generation
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