|
| Status |
Public on Apr 11, 2013 |
| Title |
Serum_healthy_6 |
| Sample type |
RNA |
| |
|
| Source name |
Serum
|
| Organism |
Felis catus |
| Characteristics |
condition: healthy
|
| Treatment protocol |
Blood was collected by venipuncture in serum tubes and left to clot for 20 min. The blood was centrifuged at 600g for 5 min and the serum was stored at -80°C until RNA extraction.
|
| Growth protocol |
All cats were client-owned adult animals (Age 6 -14 years) that were presented as patients or for health checks to the clinic of Small Animal Veterinary Medicine, LMU University of Munich, Germany
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from 16 serum samples using Qiazol® Reagent (Qiagen, Hilden Germany) and the miRNeasy Mini Kit (QIAGEN, Valencia, CA, USA) according to the supplementary protocol “Purification of total RNA, including small RNAs, from serum or plasma". Quantity and purity of RNA were measured with a NanoDrop 1000 (PEQLAB Biotechnologie GMBH, Erlangen, Germany).
|
| Label |
Cy3
|
| Label protocol |
Labeling of miRNAs with Cy3-CTP was performed with the miRNA Complete Labeling and Hybridization Kit (Cat.-No. 5190-1934; Agilent Technologies) according the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Microarray analysis was performed using custom-made Agilent miRNA microarrays. Cy3-labeled miRNA was hybridized to the microarrays according the manufacturer’s instructions (User Manual G4170-90011_miRNA_complete_2.3).
|
| Scan protocol |
Hybridized and washed slides were scanned at 2 µm resolution with an Agilent DNA Microarray Scanner (G2505C, Agilent Technologies).
|
| Description |
miRNA expression profile in the serum of a healthy cat
|
| Data processing |
Image processing was performed with Feature Extraction Software 10.7.3.1 (Agilent Technologies).
The gProcessedSignal column contains the original values from the FE software. For further statistical analysis gProcessedSignals were 1) filtered based on “Is well above background” flags (A feature was listed as detectable if > 50% of the signals in at least one of the two measured groups were above detection limit, and a probe was considered as detectable if ≥ 50% of the replicated features were detectable); 2) detectable probes were summarized (calculation of mean signal intensities for each probe); and 3) normalized with the BioConductor package vsn.
|
| |
|
| Submission date |
Apr 11, 2012 |
| Last update date |
Apr 11, 2013 |
| Contact name |
Karin Weber |
| E-mail(s) |
karin.weber@lmu.de
|
| Organization name |
Zentrum für klinische Tiermedizin
|
| Department |
Medizinische Kleintierklinik
|
| Street address |
Veterinärstr. 13
|
| City |
Munich |
| ZIP/Postal code |
80539 |
| Country |
Germany |
| |
|
| Platform ID |
GPL15426 |
| Series (1) |
| GSE37177 |
miRNA expression profiles in the serum of diabetic cats |
|
