|
Status |
Public on Jun 04, 2014 |
Title |
N487T4 |
Sample type |
RNA |
|
|
Source name |
Cord blood_#487_CD4 Tcell_CCR4
|
Organism |
Homo sapiens |
Characteristics |
donor id: #487 gender: male tissue: cord blood cell type: CD4 T cells cell subset: CCR4
|
Treatment protocol |
Activaiton with antiCD3+antiCD28
|
Growth protocol |
Culture for 24h in RPMI with 10%FCS
|
Extracted molecule |
total RNA |
Extraction protocol |
Cell samples were lysed using SuperAmp Lysis Buffer following manufacturer’s instructions. SuperAmplification was performed according to Miltenyi Biotec’s undisclosed protocol. Briefly, the amplification is based on a global PCR protocol of mRNA-derived cDNA. mRNA was isolated via magnetic bead technology.
|
Label |
Cy3
|
Label protocol |
250 ng of each of the amplified cDNAs were used as template for Cy3 labeling which was performed according to Miltenyi Biotec’s undisclosed protocol.
|
|
|
Hybridization protocol |
Cy3 labeled cDNAs were hybridized overnight (17 hours, 65°C) to an Agilent Whole Human Genome Oligo Microarrays (8x60K) using Agilent’s recommended hybridization chamber and oven. Finally, microarrays were washed once with wash buffer 1 for 1 min at RT followed by a second wash with wash buffer 2 for 1 min at 37°C and a final wash step with acetonitrile for 30 sec at RT.
|
Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent’s Microarray Scanner System (G2505C, Agilent Technologies, Palo Alto, USA).
|
Description |
39
|
Data processing |
The Agilent Feature Extraction Software (FES 10.7.3.1) was used to read out and process the microarray image files.
|
|
|
Submission date |
Sep 24, 2013 |
Last update date |
Jun 04, 2014 |
Contact name |
Richard Lo-Man |
E-mail(s) |
richard.lo-man@pasteur.fr
|
Organization name |
Institut Pasteur
|
Lab |
Neonatal Immunity group
|
Street address |
25-28 Rue du Docteur Roux
|
City |
Paris |
ZIP/Postal code |
75015 |
Country |
France |
|
|
Platform ID |
GPL16699 |
Series (1) |
GSE51133 |
Analysis of cord blood CD4T cells |
|