|
| Status |
Public on Jul 01, 2022 |
| Title |
bone metastais 3 |
| Sample type |
RNA |
| |
|
| Source name |
bone metastais 3
|
| Organism |
Homo sapiens |
| Characteristics |
disease: prostate cancer
gleason: Gleason 3+4
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA Extraction was perforemd using TRIZOL following the manufacturer's recommendations.
|
| 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 RNAeasy 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 Microarrays (8*60K) 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 |
Arrays were scanned using an Agilent Scanner (G2505C) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 5 um . Dye channel was set to Green and Green PMT is set to 100%.
|
| Description |
mirna expression profile
|
| Data processing |
The generated image files were analyzed using Agilent Feature Extraction Software (v 11.0.1.1). Using GemeSpring v12.1 software(Agilent) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Aug 05, 2020 |
| Last update date |
Jul 01, 2022 |
| Contact name |
Jing Hu |
| E-mail(s) |
waterfall_1988@aliyun.com
|
| Organization name |
Shandong University
|
| Department |
Department of Pathology
|
| Street address |
Wenhua xi road
|
| City |
Jinan |
| ZIP/Postal code |
250012 |
| Country |
China |
| |
|
| Platform ID |
GPL25134 |
| Series (1) |
| GSE155732 |
miRNAs expression of Gleason 3+4 Prostate Cancer |
|
