|
| Status |
Public on Apr 04, 2013 |
| Title |
Serum_control_pooled, rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Venous blood, control
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: blood serum
disease status: control
|
| Treatment protocol |
The whole blood was separated into serum and cellular fractions by centrifugation at 4,000 rpm for 10 min, followed by 12000 rpm for 15 min to completely remove cell debris. To detect the generalizable signatures of serum miRNAs for MMD, we pooled serum samples from 10 patients and 10 controls.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol Reagent (Invitrogen, Carlsbad, CA) was used for serum denaturizing and the Qiagen miRNeasy Mini kit (Qiagen, Valencia, CA) for RNA collection and purification according to the manufacturer's protocol.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 100 ng RNA using Agilent's miRNA Complete Labeling and Hyb Kit (p/n 5190-0456) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
Agilent Human 8 x 60K miRNA Array was performed on the two pooled samples. Hybridization was carried out according to the instructions of the Agilent's miRNA Complete Labeling and Hyb Kit (p/n 5190-0456).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Serum samples from 10 controls were pooled into a single control sample.
|
| Data processing |
Data were acquired using the Agilent G2565BA Microarray Scanner System and Agilent Feature Extraction Software (Scan resolution 5μm, PMT 100%). Probe intensities were normalized using GeneSpring 12.0 through percentile shift normalization. Differentially expressed miRNAs were identified through Fold Change filtering (Fold change >2).
|
| |
|
| Submission date |
Apr 03, 2013 |
| Last update date |
Apr 04, 2013 |
| Contact name |
Dongwei Dai |
| E-mail(s) |
lsykxy@126.com
|
| Organization name |
Changhai Hospital
|
| Street address |
168 Changhai Road
|
| City |
Shanghai |
| ZIP/Postal code |
200433 |
| Country |
China |
| |
|
| Platform ID |
GPL15159 |
| Series (1) |
| GSE45737 |
Determination of the serum miRNA expression profile in human moyamoya disease (MMD) |
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