|
| Status |
Public on Dec 26, 2020 |
| Title |
non-rejection_plasma_replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
non-rejection plasma
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: plasma
gender: male
|
| Treatment protocol |
Freshly drawn blood was collected on the 7thday after OLT in ethylenediaminetetraacetic acid (EDTA)-treated tubes. Within 30 minutes, the tubes were centrifuged at 820g for 10 min. Then, 1-ml aliquots of plasma were transferred to 1.5-ml tubes and centrifuged at 16,000g for 10 min to pellet any remaining cellular debris. Liver grafts of each animal were obtained and conserved in liquid nitrogen until use.
|
| Growth protocol |
Inbred male Lewis and BN rats weighing 200-250g were maintained in laminar flow cages in a specific pathogen-free animal facility with a standard diet and water.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from plasma samples with the mirVana PARIS miRNA Isolation Kit (Ambion, Austin, TX) and from tissue samples with the mirVana miRNA Isolation Kit (Ambion). RNA concentrations were determined with a NanoDrop 1000 Spectrophotometer (NanoDrop Technologies, Waltham, MA)
|
| Label |
Cy3
|
| Label protocol |
RNA was labeled with Cy3 according to the protocols of the Agilent microRNA microarray system
|
| |
|
| Hybridization protocol |
Each slide was hybridized with 100 ng Cy3-labeled RNA with the miRNA Complete Labeling and Hyb Kit (Agilent Technologies, Santa Clara, CA) in a hybridization oven (Agilent Technologies) at 55°C and 20 rpm for 20 hours according to the manufacturer’s instructions.
|
| Scan protocol |
Microarray slides were scanned by an XDR Scan (PMT100, PMT5) according to the protocols of the Agilent microRNA microarray system
|
| Description |
XM37plasma
|
| Data processing |
The microarray image data were converted into spot intensity values with Feature Extraction Software Rev. 9.5.3 (Agilent Technologies). The signal minus background values were exported directly into GeneSpring GX10 software (Agilent Technologies), and the raw data were normalized by the Quantile algorithm.
|
| |
|
| Submission date |
Mar 26, 2012 |
| Last update date |
Dec 26, 2020 |
| Contact name |
jie hu |
| Organization name |
Zhongshan hospital, Fudan University
|
| Street address |
180 Fenglin Road
|
| City |
Shanghai |
| ZIP/Postal code |
200032 |
| Country |
China |
| |
|
| Platform ID |
GPL10906 |
| Series (1) |
| GSE36798 |
Development of microRNA signatures for acute rejection after liver transplantation using rat model |
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