|
| Status |
Public on Oct 10, 2021 |
| Title |
Lung, WA (SAMPLE 5) |
| Sample type |
RNA |
| |
|
| Source name |
Lung, Test1
|
| Organism |
Mus musculus |
| Characteristics |
genotype: K18-hACE2 transgenic
tissue: Whole lung homogenate
infection: SARS-CoV-2
activation agent: WA infection
time: dpi 3 after WA infection
group: Test1
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using QIAgen RNeasy Plus Mini Kit
|
| Label |
SYBR Green
|
| Label protocol |
Reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). Quantitative real-time PCR were performed in Stratagene MX3000p using RT² Profiler™ PCR Array Mouse Hypoxia Signaling Pathway kit (QIAgen) following the Manufacturer’s instructions. The cycling conditions are: hold for 10 min at 95°C, followed by 40 cycles of 15 s at 95°C and 60 s at 60°C.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Data processing |
The normalization and all the data analysis were performed according to the manufacturers instructions using their web-based software package: https://dataanalysis.qiagen.com/pcr/arrayanalysis.php?wuid=b4d56157-c14b-4fd9-a99c-8ddecd502e64&logindata=&customerdata=&customeremail=&platform=catalogArray&format=X
Normalization was performed using GUSB as the internal housekeeping gene.
Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)]
The web-based software package automatically performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data.
non-normalized_data.txt reports raw signals
normalized_data.txt reports normalized signal (against housekeeping gene).
Fold_Change.txt file reports test/control (i.e., NY/Naive) ratios.
|
| |
|
| Submission date |
Sep 07, 2021 |
| Last update date |
Oct 10, 2021 |
| Contact name |
Hang Xie |
| E-mail(s) |
Hang.Xie@fda.hhs.gov
|
| Organization name |
US Food and Drug Administration, Center for Biologics Evaluation and Research
|
| Street address |
10993 New Hampshire Ave WO 52/72 Rm 1324
|
| City |
Silver Spring |
| State/province |
Maryland |
| ZIP/Postal code |
20993 |
| Country |
USA |
| |
|
| Platform ID |
GPL30599 |
| Series (1) |
| GSE183621 |
Real-time quantitative PCR analysis of mouse lungs after exposure to SARS-CoV-2 and variants of concern (VOC) |
|
