|
| Status |
Public on Feb 22, 2017 |
| Title |
CD8+T cell activated TET2cKO rep2 |
| Sample type |
RNA |
| |
|
| Source name |
CD8+ T cell, activated in vitro, 3d
|
| Organism |
Mus musculus |
| Characteristics |
genotype: TET2cKO
|
| Treatment protocol |
N/A
|
| Growth protocol |
WT or TET2-deficient naive CD8+ T cells were cultured on plate-bound anti-CD3 (1ug/ml) and anti-CD28 (5ug/ml) with soluble hIL-2 (100U/ml) in complete IMDM for 3 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using RNeasy MIni kit (Qiagen) according to manufacturer's instructions. Total RNA was assessed by UV absorbance and all had 260/280 ratios above 1.7. Additionally, samples were run on the Agilent RNA 6000 Nano chip and all had RINs above 8.6.
|
| Label |
biotin
|
| Label protocol |
Sense-strand single-stranded cDNA was prepared from 200ng of total RNA using the Ambion WT Expression Kit (PN: 4411974). Both cRNA and cDNA were assessed using UV absorbance. 5.5ug of single-stranded cDNA was fragmented and labeled using the Affymetrix GeneChip WT Terminal Labeling kit (PN: 900671).
|
| |
|
| Hybridization protocol |
Affymetrix GeneChip Hybridization Wash and Stain Kit (PN: 900720) protocol was used for all hybridization, wash and staining steps. Fragmented and labeled cDNA (130ul at 25ng/uL) were hybridized for 17hrs at 45C to Affymetrix GeneChip Mouse Gene 2.0ST Arrays (PN: 902118).
|
| Scan protocol |
Chips were washed and stained on an Affymetrix Genechip Wash Station 450 using the FS450_0002 protocol and scanned on the Affymetrix GeneChip Scanner 3000. Cel files were generated for analysis using Affymetrix GeneChip Command Console v3.2.
|
| Data processing |
Affymetrix GeneChip Command Console v3.2.
|
| |
|
| Submission date |
Feb 21, 2017 |
| Last update date |
Feb 22, 2017 |
| Contact name |
Shannon Carty |
| E-mail(s) |
scarty@med.umich.edu
|
| Organization name |
University of Michigan
|
| Street address |
109 Zina Pitcher Place
|
| City |
Ann Arbor |
| State/province |
MI |
| ZIP/Postal code |
48109 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE95147 |
Gene expression data from WT and TET2cKO CD8+ T cells activated in vitro |
|
