|
| Status |
Public on Apr 16, 2013 |
| Title |
Sorted KLRG-1 Lo memory precursor Db-GP33 specific CD8 T cells from infected mice, Replicate 3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Direct ex-vivo splenocytes
|
| Organism |
Mus musculus |
| Characteristics |
days post-infection: Day 4.5 following LCMV Armstrong infection
cell type: Purified CD8 T cells
antigen specificity: GP33-41 epitope of LCMV Glycoprotein
strain: C57BL/6
|
| Growth protocol |
GP33-41 specific P14 cells were isolated and sorted from mice
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using miRNeasy kit (Qiagen) following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Total of 1ug of RNA isolated from FACS sorted CD8 T cells was 3' end labeled with Hy3 following the Exiqon miRCURY LNA Labeling Kit protocol. In parallel, 1ug of Common Reference Design pool RNA was labeled with Cy5 for control. The Common Reference Pool Design control was generated per Exiqon's guidelines by pooling equal amounts of all the experimental RNA samples.
|
| |
|
| Channel 2 |
| Source name |
Pooled total RNA from all the experimental samples
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 4-6 weeks
Sex: Female
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using miRNeasy kit (Qiagen) following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Total of 1ug of RNA isolated from FACS sorted CD8 T cells was 3' end labeled with Hy3 following the Exiqon miRCURY LNA Labeling Kit protocol. In parallel, 1ug of Common Reference Design pool RNA was labeled with Cy5 for control. The Common Reference Pool Design control was generated per Exiqon's guidelines by pooling equal amounts of all the experimental RNA samples.
|
| |
|
| |
| Hybridization protocol |
Hybridizations were performed using Exiqon's protocol. Briefly, the labeled samples were hybridized for 16-18 hours at 60C and then washed sequentially in 3 specially designed buffers at recommended temperatures and conditions.
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner.
Images were quantified using Agilent Feature Extraction Software (version A.7.5).
|
| Description |
Pooled total RNA - called the Common Reference Design Pool
|
| Data processing |
LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used. 156 probes that lit up for our mouse samples, the rest of the values were either missing in some samples or the miRNA was not designed for mice.
|
| |
|
| Submission date |
Apr 15, 2013 |
| Last update date |
Apr 16, 2013 |
| Contact name |
Surojit Sarkar |
| Organization name |
University of Washington School of Medicine
|
| Department |
Department of Pediatrics and Laboratory Medicine
|
| Lab |
Laboratory of T Cell Immunity to Pathogens and Cancer
|
| Street address |
1100 Olive Way, Suite 100
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98101 |
| Country |
USA |
| |
|
| Platform ID |
GPL7724 |
| Series (1) |
| GSE46052 |
miR-17~92 Regulates Effector and Memory CD8 T cell Fates by Modulating Proliferation in Response to Infection |
|
