SurePrint G3 Human Gene Expression 8x60K v2 Microarray Kit

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	In the discovery state, by comparing the relative expression of serum miRNAs, up- or down-regulated miRNAs were selected. We identified the expression of 2,550 miRNAs using the SurePrint G3 Human miRNA Microarray, Release 21.0 (Agilent, Santa Clara, CA, USA). Total RNA (100 ng) was treated with phosphatase and incubated at 37°C for 30 minutes. Dimethyl sulfoxide was added to the dephosphorylated RNA, followed by the application of heat and ice. Then, the dephosphorylated RNA was incubated at 16°C for 2 hours for assembly of the labeling reaction. The labeled RNA was desalted using a spin column. The desalted labeled RNA was dried with a vacuum concentrator at 45°C to 55°C for approximately 2 to 3 hours, followed by hybridization to the Agilent miRNA expression microarray at 55°C for 20 hours. Arrays were scanned using the Agilent Technologies G2600D SG12494263 microarray platform.

Protocol tips

In the discovery state, by comparing the relative expression of serum miRNAs, up- or down-regulated miRNAs were selected. We identified the expression of 2,550 miRNAs using the SurePrint G3 Human miRNA Microarray, Release 21.0 (Agilent, Santa Clara, CA, USA). Total RNA (100 ng) was treated with phosphatase and incubated at 37°C for 30 minutes. Dimethyl sulfoxide was added to the dephosphorylated RNA, followed by the application of heat and ice. Then, the dephosphorylated RNA was incubated at 16°C for 2 hours for assembly of the labeling reaction. The labeled RNA was desalted using a spin column. The desalted labeled RNA was dried with a vacuum concentrator at 45°C to 55°C for approximately 2 to 3 hours, followed by hybridization to the Agilent miRNA expression microarray at 55°C for 20 hours. Arrays were scanned using the Agilent Technologies G2600D SG12494263 microarray platform.

Publication protocol

In the discovery state, by comparing the relative expression of serum miRNAs, up- or down-regulated miRNAs were selected. We identified the expression of 2,550 miRNAs using the SurePrint G3 Human miRNA Microarray, Release 21.0 (Agilent, Santa Clara, CA, USA). Total RNA (100 ng) was treated with phosphatase and incubated at 37°C for 30 minutes. Dimethyl sulfoxide was added to the dephosphorylated RNA, followed by the application of heat and ice. Then, the dephosphorylated RNA was incubated at 16°C for 2 hours for assembly of the labeling reaction. The labeled RNA was desalted using a spin column. The desalted labeled RNA was dried with a vacuum concentrator at 45°C to 55°C for approximately 2 to 3 hours, followed by hybridization to the Agilent miRNA expression microarray at 55°C for 20 hours. Arrays were scanned using the Agilent Technologies G2600D SG12494263 microarray platform.

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