|
| Status |
Public on Nov 01, 2018 |
| Title |
EMca_NORMAL_46 |
| Sample type |
RNA |
| |
|
| Source name |
Normal_endometrial_tissue
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: normal endometrial tissue
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA, including miRNA, was extracted using a QIAzol Lysis reagent (Qiagen, Valencia, CA, USA) and a miRNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by miRNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent human miRNA V2 Oligo Microarray Kit (G4470B) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
The arrays were scanned with an Agilent microarray scanner (Agilent Technologies) using high dynamic range settings as specified by the manufacturer.
|
| Description |
normal endometrial tissues_46
|
| Data processing |
Microarray results were extracted using Agilent Feature Extraction software Ver. 9.5.3.1 (Agilent Technologies) and analyzed using Gene Spring GX 7.3.1 software (Agilent Technologies) to obtain gene expression ratios. Ratio of intensities values above or less than 2.0 were considered significant for upregulated or downregulated genes.
|
| |
|
| Submission date |
Nov 16, 2010 |
| Last update date |
Nov 01, 2018 |
| Contact name |
ERI HIROKI |
| Organization name |
Tohoku University
|
| Department |
Obstetrics and Gynecology
|
| Street address |
1-1 Seiryo-Machi, Aoba-ku
|
| City |
Sendai |
| ZIP/Postal code |
980-8574 |
| Country |
Japan |
| |
|
| Platform ID |
GPL7731 |
| Series (1) |
| GSE25405 |
EMca: serous, endometrioid, normal (miRNA) |
|
