|
| Status |
Public on Jul 06, 2018 |
| Title |
ISP WT_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
ISP T-cells from WT mouse
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
cell type: Immature Single Positive T-cells
genotype: WT
|
| Treatment protocol |
Sorted cells were lysed with lysis buffer from Rneasy Micro kit from Qiagen.
|
| Growth protocol |
WT and Rptor-/- cell populations were purified from mouse thymus by fluorescence activated cell sorting (FACS) based on the following cell surface markers: DN3a thymocytes (CD4-CD8-TCRγδ-TCRβ-CD25hiCd44-CD28-); ISP T-cells (CD4-CD8+TCRβ-); gd T-cells (CD4-CD8-TCRb-NK1-1-Gr1-).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA samples were prepared according to the prototol of Rneasy Micro kit from Qiagen,
|
| Label |
biotin
|
| Label protocol |
Twelve (12) nanograms of total RNA were converted into biotinylated cDNA using the NuGEN WTA Pico v2 protocol and the NuGEN Encore Biotin module
|
| |
|
| Hybridization protocol |
Six micrograms of biotinylated cDNA was fragmented and then hybridized for 16 hr at 45C on an Affymetrix Mouse Gene 2.0 ST GeneChip. Following hybridization, GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the AffymetrixGeneChip Scanner 3000 7G.
|
| Data processing |
Probe signals were quantile normalized and summarized by the RMA algorithm using Partek Genomics Suite software v6.6
|
| |
|
| Submission date |
Sep 06, 2016 |
| Last update date |
Jul 07, 2018 |
| Contact name |
Geoffrey Neale |
| E-mail(s) |
geoffrey.neale@stjude.org
|
| Organization name |
St Jude Childrens Research Hospital
|
| Department |
Hartwell Center
|
| Street address |
262 Danny Thomas Place
|
| City |
Memphis |
| State/province |
TN |
| ZIP/Postal code |
38105 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE86495 |
Metabolic signaling directs the reciprocal lineage decisions of αβ and γδ T cells |
|
