|
| Status |
Public on Mar 18, 2010 |
| Title |
CD62Llo PSGL1lo, technical replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
TCRb+ CD4+ B220- CD44+ splenocytes isolated from aged mice
|
| Organism |
Mus musculus |
| Characteristics |
tissue: spleen
genotype: MRL/Faslpr
age: 16-24 weeks
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using the Qiagen RNAeasy kit according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Single stranded cDNA is synthesized by reverse transcription using the poly (A) RNA present in the starting total RNA sample. Single stranded cDNA is then converted into double stranded cDNA and purified using the Affymetrix Cleanup Module.
An in vitro transcription (IVT) reaction is then carried out overnight in the presence of biotinylated UTP and CTP to produce biotin-labeled cRNA from the double stranded cDNA. The resulting cRNA is fragmented in the presence of heat and Mg+2 before hybridization to the test array.
|
| |
|
| Hybridization protocol |
Fragmented cRNA was hybridized to the Affymetrix 430 2.0 mouse array for 16 hours at 45C. The array is then washed and stained with streptavidin-phycoerythrin (SAPE) using the fluidics station.
|
| Scan protocol |
GeneChips were scanned using GeneChip Scanner 3000.
|
| Description |
Gene expression data from CD62Llo PSGL1lo effector (CD44hi) CD4 T cells
|
| Data processing |
The normalization algorithm used was MAS5. Additional normalization using a GeneSpring GX Analysis Platform (Agilent Technologies) used all default settings. Genes that were differentially expressed (p value ≤ 0.05) among CD62Llo PSGL1lo, CD62Llo PSGL1hi, CD62Lhi PSGL1hi, and naïve subsets (n = 3) were identified using 2-way ANOVA.
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| |
|
| Submission date |
Mar 10, 2010 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Amanda Poholek |
| E-mail(s) |
[email protected]
|
| Organization name |
Yale University
|
| Street address |
300 Cedar St. TAC S530
|
| City |
New Haven |
| State/province |
CT |
| ZIP/Postal code |
06519 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE20735 |
Expression data from effector CD4 T cells isolated from MRL/Faslpr mice. |
|
| Relations |
| Reanalyzed by |
GSE119085 |
