|
| Status |
Public on Mar 12, 2012 |
| Title |
RNA isolated from PBMCs (IgAN_5) |
| Sample type |
RNA |
| |
|
| Source name |
IgAN Patient with a Nomal Renal Function
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: IgAN
status: normal renal function
creatinine clearance: >90 ml/min/1.73 m2 body surface area (evaluated by Cockcroft-Gault formula)
serum creatinine: <1.2 mg/dl
clinical follow up: At least 7 years
|
| Treatment protocol |
Blood sample was drawn into Lithium Heparin blood collection tube
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNAs were extracted using Qiagen miRNeasy kit according to the manufacturer's instructions. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
600ng of total RNA from each sample was dephosphorylated with Calf Intestinal Alkaline Phosphatase (CIP) at 37ºC for 30’ following a denaturalization and then a ligation for 2h at 16ºC using Agilent miRNA Complete Labeling and Hyb Kit (Agilent Technologies; Palo Alto, CA). In this step a molecule of Cyanine3-pCp is incorporated to the 3’ end of RNA molecules.
|
| |
|
| Hybridization protocol |
Cy3-labeled RNA from each sample was hybridized to an Agilent Human miRNA Version 2 Microarray using the miRNA v1 Protocol
|
| Scan protocol |
The hybridized array was then washed and scanned according to Agilent specifications and data was extracted from the scanned image using Feature Extraction version 9.5.1.1. (Agilent Technologies).
|
| Description |
miRNA expression in IgAN patient
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5.1.1 (Agilent) using default parameters (Grid: 019118_D_20080214) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Nov 22, 2010 |
| Last update date |
Mar 12, 2012 |
| Contact name |
Sharon Natasha Cox |
| E-mail(s) |
sharonnatasha.cox@gmail.com
|
| Organization name |
Schena Foundation
|
| Department |
Renal Lab
|
| Street address |
Strada Provinciale Valenzano-Casamassima-Km3
|
| City |
Valenzano-Bari |
| ZIP/Postal code |
70010 |
| Country |
Italy |
| |
|
| Platform ID |
GPL7731 |
| Series (1) |
| GSE25590 |
miR-148b modulates the C1GALT1 expression explaining the abnormal glycosylation process in IgA Nephropathy |
|
