|
| Status |
Public on Jul 26, 2018 |
| Title |
Neonatal Clone 2_miRNA_ISS-cultured |
| Sample type |
RNA |
| |
|
| Source name |
Primary cardiac tissue
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Cardiovascular progenitor cell
culture condition: Biocell (BioServe Space Technologies) - ISS
duration: 12 days
|
| Growth protocol |
CPCs were flown to the ISS where they were cultured for 12 days and fixed in RNA protect. Clone-, patient-, and passage-matched controls were seeded and grown on the ground for 12 days as a control.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIzol and alcohol-based precipitation
|
| Label |
SYBR Green
|
| Label protocol |
PCR assays were performed using human development and differentiation microarray plates (MIHS-103ZA; Qiagen, Valencia, CA) following the Manufacturer’s instructions. Reverse transcription was performed from 500 ng total RNA using the miScript SYBR Green PCR master mix and 10X universal primer mix (Qiagen, Valencia, CA). Quantitative real-time PCR were performed (iQ5 thermal cycler, BioRad) with 40 cycles at 94 oC for 15 seconds, 55 oC for 30 secons, and 70 oC for 30 seconds.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
ISS-cultured
SAMPLE 4
|
| Data processing |
Normalization and comparative Ct method analysis per clone was perofmred using Qiagen's web-based software package: http://www.qiagen.com/ us/shop/genes-and-pathways/data-analysis-center-overview-page/
For normalization, we used the average of all available housekeeping genes included on the microarray plates
Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)].
Fold changes per clone were exported to Prism for subsequent analysis. Fold Change worksheet reports test/control (i.e., ISS-cultured CPCs/ground CPCs) ratios.
|
| |
|
| Submission date |
Feb 13, 2018 |
| Last update date |
Jul 26, 2018 |
| Contact name |
Jonathan Baio |
| E-mail(s) |
jbaio@llu.edu
|
| Organization name |
Loma Linda University School of Medicine
|
| Department |
Pathology and Human Anatomy
|
| Lab |
Kearns-Jonker
|
| Street address |
24941 Stewart St, Room 326
|
| City |
Loma Linda |
| State/province |
CA |
| ZIP/Postal code |
92354 |
| Country |
USA |
| |
|
| Platform ID |
GPL24607 |
| Series (2) |
| GSE110561 |
Real-time quantitative PCR analysis of human cardiovascular progenitor cells flown aboard the International Space Station [miRNA] |
| GSE110563 |
Real-time quantitative PCR analysis of human cardiovascular progenitor cells flown aboard the International Space Station |
|
