|
| Status |
Public on Mar 01, 2014 |
| Title |
P_0C1 |
| Sample type |
RNA |
| |
|
| Source name |
PBMC Control 0h
|
| Organism |
Mus musculus |
| Characteristics |
background strain: C3H/HeJ
gender: Male
tissue: PBMC
|
| Treatment protocol |
Animals were anesthetized and SEB toxin (treatment) or saline (control) was administered i.n. (5 μg SEB/ 50 μl) followed by and i.p. (2 μg SEB/200 μ-l) dose 2h later. PBMC, spleen, lung, liver, kidney, and heart were harvest for tRNA purification from SEB and saline control groups at 0, 2.75, 5 and 24 h after treatment .
|
| Growth protocol |
Male C3H/HeJ, weighing ~ 20 g each (7-10 weeks old), were housed in conventional micro isolator cages. Food and water were freely available at all times.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
PBMC were isolated by gradient (Nycoprep) from blood collected in EDTA-treated tubes and lysed in RLT buffer. Tissues were thinly sliced and preserved in RNAlater reagent at -80 C. tRNA was obtained using RNeasy kit (Qiagen) following manufacturer protocols. tRNA concentration was measure by nanodrop and tRNA quality was evaluated by Bioanalyzer.
|
| Label |
Biotin
|
| Label protocol |
Affymetrix GeneChip Expression 3'-Amplification One-Cycle Target Labeling and Control Kit was used for the synthesis of cDNA and cRNA and biotin-labeled cRNA. 2 μg of tRNA was utilized as starting material following manufacturer protocol.
|
| |
|
| Hybridization protocol |
20 μg of fragmented biotin-labeled cRNA was used in hybridization cocktails on 430 2.0 Mouse Genome Arrays (Affymetrix) following manufacturer protocols.
|
| Scan protocol |
Affymetrix 430 2.0 array chips were scanned using the Affymetrix GeneChip Scanner 7G.
|
| Description |
RLD_Oct08_PBMC_0h-1
|
| Data processing |
Data were generated using the Mas 5.0 algorithm in the Affymetrix Expression Console software
|
| |
|
| Submission date |
Nov 18, 2013 |
| Last update date |
Dec 12, 2023 |
| Contact name |
Gabriela A Ferreyra |
| E-mail(s) |
gferreyra@cc.nih.gov
|
| Phone |
301-435-2317
|
| Organization name |
Clinical Center
|
| Department |
CCMD
|
| Street address |
9000 Rockville Pike - Building 10 room 4D03
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE52474 |
Late Multiple Organ Surge in Interferon-regulated Target Genes Characterizes Staphylococcal Enterotoxin B Lethality |
|
