|
| Status |
Public on Oct 30, 2015 |
| Title |
genomic DNA from in vitro activated naive B cells [BN287_P2_Hpos] |
| Sample type |
genomic |
| |
|
| Source name |
in vitro activation of naive B cell from normal blood
|
| Organism |
Homo sapiens |
| Characteristics |
patient id: BN287
subpopulation labeling: DAPI neg CFSE low CD38 low Hoescht + (S/G2/M)
day of isolation: D6
|
| Treatment protocol |
Stimulation cocktail 1 (2,6 µg/ml Fab'2 anti-Ig, 100 ng/ml CD40 Ligand, 1 µg/ml CpG ODN 2006 and 50 U/ml IL2) for 4 days then differentiation cocktail 2 (50 U/ml IL2, 5 ng/ml IL4, 10 ng/ml IL10) for 2 days (Le Gallou et al., 2012)
|
| Growth protocol |
Naive B cells from healthy donors were purified using magnetic cell separation (Miltenyi Biotech), CFSE-labeled and cultured at 7.5.105 cells/ml for 4 days with cocktail 1. Activated B cells were then washed and further cultured at 4.105 with cocktail 2 (Le Gallou et al., 2012)
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
DNA extracted using the DNA/RNA minikit (Qiagen) after cell sorting (FACS Aria, BD Biosciences)
|
| Label |
Cy5 and Cy3
|
| Label protocol |
Standard Illumina Protocol
|
| |
|
| Hybridization protocol |
Bisulphite converted DNA was amplified, fragmented and hybridised to Illumina Infinium Human Methylation450 Beadchip using the standard Illumina protocol
|
| Scan protocol |
Arrays were imaged using the iScan reader using standard recommended Illumina scanner setting
|
| Data processing |
Using the minfi R package available through Bioconductor
|
| |
|
| Submission date |
Aug 28, 2015 |
| Last update date |
Oct 30, 2015 |
| Contact name |
Gilles Salbert |
| E-mail(s) |
gilles.salbert@univ-rennes1.fr
|
| Phone |
+33 223 236 625
|
| Organization name |
University of Rennes 1
|
| Department |
Biology
|
| Lab |
UMR6290 CNRS
|
| Street address |
Avenue du Général Leclerc
|
| City |
Rennes |
| ZIP/Postal code |
35042 |
| Country |
France |
| |
|
| Platform ID |
GPL13534 |
| Series (2) |
| GSE72495 |
Cell cycle-dependent reconfiguration of the DNA (hydroxy) methylome during terminal differentiation of human B cells into plasma cells [methylation array] |
| GSE72498 |
Cell cycle-dependent reconfiguration of the DNA (hydroxy) methylome during terminal differentiation of human B cells into plasma cells |
|
