|
| Status |
Public on Jun 13, 2018 |
| Title |
PBMC of non-infected dogs_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
PBMC of healthy and non-infected dogs
|
| Organism |
Canis lupus familiaris |
| Characteristics |
cell type: PBMC
infected with: none
|
| Extracted molecule |
total RNA |
| Extraction protocol |
PBMC were isolated by gradient Histopaque® 1077 (Sigma-Aldrich, St. Louis, MO, USA) following the manufacturer's instructions. Total RNA was extracted from 5x106 cells with a miRVana™ miRNA Isolation Kit (Ambion, Austin, TX, USA) according to the manufacturer’s instructions. RNA samples were immediately stored at -80ºC. Quantification was performed using NanoDrop (ND-2000 spectrophotometer, Thermo Scientific, Wilmington, DE, USA) and quality of samples was assessed using a Capillary electrophoresis (Bioanalyzer 2100 Agilent, Santa Clara, CA, USA).
|
| Label |
biotin
|
| Label protocol |
MicroRNA profiles were obtained from 250 ng/sample of total RNA (RNA Integrity Number over than 7) using the FlashTag™ Biotin HSR RNA Labeling Kit, and the Affymetrix® miRNA 4.1 Array strip (Affymetrix, Santa Clara, Califórnia, EUA), according to the manufacturer’s instructions. A recommended ELOSA quality control assay was run for all samples.
|
| |
|
| Hybridization protocol |
For hybridization of samples to the strips was carried on at 48ºC for 20h using the GeneAtlas® Hybridization, Wash, and Stain Kit for miRNA Array Strips. Following this period, strips were processed and scanned using the GeneAtlas® System (Affymetrix).
|
| Scan protocol |
After the array strip are registered in the GeneAtlas Software and the target hybridization steps are complete, scanning processing on the GeneAtlas System was made. Transfer of the strip assembly into the imaging station's drawer starts the imaging process.
|
| Description |
miRNA profile expression in PBMC of non-infected dogs
|
| Data processing |
Raw intensity values were background corrected, log2 transformed and then quantile normalized by the software Expression Console (Affymetrix) using the Robust Multi-array Average (RMA) algorithm.
|
| |
|
| Submission date |
Oct 20, 2017 |
| Last update date |
Jun 13, 2018 |
| Contact name |
Jaqueline Poleto Bragato |
| E-mail(s) |
jaquelinepoleto@hotmail.com
|
| Phone |
+5518996237632
|
| Organization name |
Universidade Estadual Paulista Júlio de Mesquita Filho - UNESP
|
| Department |
Department of animal clinic, surgery and reproduction
|
| Lab |
Immunology
|
| Street address |
Clóvis Pestana, 793
|
| City |
Araçatuba |
| State/province |
SP |
| ZIP/Postal code |
16050-680 |
| Country |
Brazil |
| |
|
| Platform ID |
GPL21572 |
| Series (1) |
| GSE105443 |
Expression data of miRNAs from PBMC of dogs infected with L. infantum |
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