|
| Status |
Public on Oct 23, 2012 |
| Title |
BM3 |
| Sample type |
RNA |
| |
|
| Source name |
MVA-HIV gag injected_bone marrow
|
| Organism |
Mus musculus |
| Characteristics |
strain background: Balb/c
injected with: modified virus Ankara (MVA)-HIV gag
time point: 5 days after intradermal injection
tissue: bone marrow
cell population: pentamer H2kD-AMQMLKETI CD8 T cells
|
| Treatment protocol |
Cells sorted by flow cytometry from fresh mouse lymph node and bone marrow cell suspensions: pentamer H2kD-AMQMLKETI (HIV-gagP24) CD8+ CD3+ T cells from draining lymph nodes (LN) and bone marrow (BM ) 5 days following intradermal injection of MVA-HIV gag, CD62L Hi CD44 int CD8+CD3+ T cells from lymph node of non-injected mice (NTc).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell samples were lysed using SuperAmp Lysis Buffer following manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
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. 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 Mouse 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 |
gene expression 5 days after intradermal injection of MVA-HIV gag
|
| 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 |
Oct 22, 2012 |
| Last update date |
Oct 23, 2012 |
| Contact name |
Helene Perrin |
| E-mail(s) |
helene.perrin0@upmc.fr
|
| Organization name |
INSERM
|
| Department |
U945
|
| Lab |
B. Combadiere
|
| Street address |
91 boulevard de l'hopital
|
| City |
Paris |
| ZIP/Postal code |
75013 |
| Country |
France |
| |
|
| Platform ID |
GPL10787 |
| Series (1) |
| GSE41736 |
Murine HIV-gagP24 CD8 T cells: bone marrow vs Lymph node |
|
