|
| Status |
Public on Apr 13, 2019 |
| Title |
LNCaP CUL4B knockdown |
| Sample type |
RNA |
| |
|
| Source name |
prostate cancer
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: prostate cancer
cell line: LNCaP
Sex: Male
genotype/variation: CUL4B-depleted
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted by Absolutely RNA miRNA Kit (Agilent p/n 400814) following the manufacturer’s instructions.
|
| 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 |
Slides were scanned immediately after washing on the Agilent Microarray Scanner using oneAgilentG3_miRNA setting for 8x60K array slides.
|
| Data processing |
The scanned images were analyzed with Agilent Feature Extraction software V.11.5 using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Apr 12, 2019 |
| Last update date |
Apr 13, 2019 |
| Contact name |
meng jiao |
| E-mail(s) |
jiaomeng851@163.com
|
| Organization name |
2nd Hospital of Shandong University
|
| Street address |
No.247, beiyuan street
|
| City |
Jinan |
| ZIP/Postal code |
250033 |
| Country |
China |
| |
|
| Platform ID |
GPL25134 |
| Series (1) |
| GSE129737 |
MicroRNA expression in CUL4B-depleted LNCaP cells and control |
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