|
| Status |
Public on Sep 05, 2009 |
| Title |
Donor 1, EBNA-3A mutant A infected LCL |
| Sample type |
RNA |
| |
|
| Source name |
human lymphoblastoid cell line (EBV transformed)
|
| Organism |
Homo sapiens |
| Characteristics |
time in culture: 6 months
cell type: lymphoblastoid
infection: EBNA-3A mutant A
|
| Treatment protocol |
1-2 x 10e5 CD19 positive B cells were infected with 1000 GRUs of EBVwt or different viral mutants and plated on lethally irradiated MRC5 feeder layer in a total volume of 200µl/well in a 96-well cluster plate. Once per week 100µl of the culture medium was replaced with fresh medium. 28-35 days p.i. proliferating B-cell cell cultures were removed from feeder layers and expanded indefinetly in suspension.
|
| Growth protocol |
Human primary B cells were isolated from adenoids, depleted of T cells by rosetting with sheep erytrocytes, and purified by Ficoll-Hypaque density-gradient centrifugation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total cellular RNA was extracted from established cell lines using the the peqGOLD Trifast kit according to the manufacturer's protocol.
|
| Label |
biotin
|
| Label protocol |
1 µg total cellular RNA was reverse transcribed and biotinylated cRNA generated according to Affymetrix protocols.
|
| |
|
| Hybridization protocol |
standard Affymetrix protocol
|
| Scan protocol |
standard Affymetrix protocol
|
| Description |
donor 1, EBNA-3A mutant A infected LCL
|
| Data processing |
The probe level data (CEL files) were processed for global normalization and generation of expression values using the robust multi-array average algorithm (RMA) of the “affy package” in the R software environment (http://www.R-project.org).
|
| |
|
| Submission date |
Sep 01, 2009 |
| Last update date |
Jun 03, 2013 |
| Contact name |
Roland Lang |
| E-mail(s) |
roland.lang@uk-erlangen.de
|
| Organization name |
University Hospital Erlangen
|
| Street address |
Wasserturmstr. 3-5
|
| City |
Erlangen |
| ZIP/Postal code |
91054 |
| Country |
Germany |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE17908 |
Differential gene expression patterns of EBV infected EBNA-3 positive and negative human B lymphocytes |
|
