|
| Status |
Public on Jul 01, 2009 |
| Title |
NKT 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
NKT 2
|
| Organism |
Mus musculus |
| Characteristics |
cell type: NKT cell
strain: C57BL/6
age: 4 weeks
tissue: 18 pooled thymi
|
| Treatment protocol |
Enrichment of HSA low mature thymocytes by anti-HSA mAb and rabbit complement; sorting of mature NKT and T cells
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRVana kit (Ambion)
|
| Label |
Hy3
|
| Label protocol |
miRCURY LNA Array power labeling kit (Exiqon, Denmark)
|
| |
|
| Channel 2 |
| Source name |
Total thymocytes
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 4 weeks
tissue: 18 pooled thymi X 3 replicates; pool of all the thymi used in the sortings; used as a common reference
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRVana kit (Ambion)
|
| Label |
Hy5
|
| Label protocol |
miRCURY LNA Array power labeling kit (Exiqon, Denmark)
|
| |
|
| |
| Hybridization protocol |
The Hy3™-labeled samples and a Hy5™-labeled reference RNA sample were mixed pair-wise and hybridized to the miRCURY™ LNA array version 11.0 (Exiqon, Denmark), which contains capture probes targeting all miRNAs for human, mouse or rat registered in the miRBASE version 12.0 at the Sanger Institute. The hybridization was performed according to the miRCURY™ LNA array manual using a Tecan HS4800 hybridization station (Tecan, Austria).
|
| Scan protocol |
Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., USA) and the image analysis was carried out using the ImaGene 8.0 software (BioDiscovery, Inc., USA).
|
| Description |
microRNA expression
|
| Data processing |
The quantified signals were background corrected (Normexp with offset value 10 – Ritchie et al., 2007) and normalized using the global Lowess (LOcally WEighted Scatterplot Smoothing) regression algorithm.
|
| |
|
| Submission date |
May 01, 2009 |
| Last update date |
Nov 19, 2010 |
| Contact name |
Maya Fedeli |
| E-mail(s) |
fedeli.maya@hsr.it
|
| Phone |
00390226434735
|
| Organization name |
San Raffaele Scientific Institute
|
| Department |
DIBIT
|
| Lab |
Experimental Immunology Unit
|
| Street address |
via Olgettina 58
|
| City |
Milan |
| State/province |
MI |
| ZIP/Postal code |
20132 |
| Country |
Italy |
| |
|
| Platform ID |
GPL7723 |
| Series (1) |
|
