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| Status |
Public on Mar 20, 2022 |
| Title |
sham-operation-3W rat bladder-rep1 |
| Sample type |
RNA |
| |
|
| Source name |
sham-operation-3 weeks rat bladder-replicate 1
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: bladder
gender: female
|
| Treatment protocol |
S group: A urethral catheter (outer diameter: 1 mm) was placed through the urethra of anesthetized female rats and ligated the proximal urethra with 4-0 wires to simulate pBOO, C group: sham operation group, urethral ligation was not performed. B group: PBOO rat treated with tolterodine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the RNasey Mini Kit (Qiagen, Hilden, Germany) following the manufacturer's recommendations. The protocol includes Five bladder specimens from each group were examined. RNA quantification and quality assurance by NanoDrop-1000 and RNA integrity and gDNA contamination test by Denaturing Agarose Gel Electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 1 ug RNA using the One-Color Quick Amp Labeling Kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy mini kit (Qiagen, Hilden, Germany). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
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| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 25x Agilent fragmentation buffer and 10x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x GEx Hybridization Buffer HI-RPM was added to the fragmentation mixture and hybridized to Agilent Whole Rat Genome Microarrays (GPL14746) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with Gene Expression Wash Buffer 1(Agilent) and 1 minute with 37°C Gene Expression Wash Buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent Microarray Scanner (G2565BA) using one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Gene expression in sham-operation-3 weeks rat bladder
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded
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| |
|
| Submission date |
Mar 20, 2019 |
| Last update date |
Mar 21, 2022 |
| Contact name |
Junyu Lai |
| E-mail(s) |
[email protected]
|
| Organization name |
Sichuan University
|
| Street address |
No. 37 Guo Xue Xiang, Chengdu, Sichuan
|
| City |
Chengdu |
| ZIP/Postal code |
610041 |
| Country |
China |
| |
|
| Platform ID |
GPL14746 |
| Series (1) |
| GSE128618 |
Activation of β-adrenoceptor signaling inhibits extracellular matrix deposition of human bladder smooth muscle cells |
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