|
| Status |
Public on Aug 30, 2017 |
| Title |
hiPS R4 (Feeder free) |
| Sample type |
RNA |
| |
|
| Source name |
hiPS R4 (Feeder free)
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
cell line: R4 iPSC line derived from MRC5 cells using lentiviral constructs separately expressing OSKM. Grown in feeder-free conditions
protocol: derived using lentiviral constructs separately expressing OSKM, grown in feeder-free conditions (vitronectin-coated plates and Essential 8 media)
passage: 20
|
| Treatment protocol |
On day 0, about 1-1.2 x 10^5 cells grown on 6-well plates were infected with a combination of lentiviruses separately encoding O,S,K and M (OSKM samples) or O, S(v), K and M (OS(v)KM samples) with 4 ug/ml polybrene. Mock- infected MRC5 cells were treated with 4 ug/ml polybrene alone. 16-18 hours later, media was replaced with EMEM + 10% FBS and changed thereafter daily. Cells were harvested on day 5 by adding 1 ml Trizol to each well.
|
| Growth protocol |
MRC5 (mock and infected) cells were maintained in Eagle's Minimal Essential Medium (EMEM, ATCC Cat # 30-2003) supplemented with 10% FBS
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with the Trizol reagent (1ml/ 6 well; ThermoFisher Cat # 15596-018) according to manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Illumina protocol from total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, cRNA were hybridized for Illumina GX HT12 Array. Slides were washed and stained in the Illumina instrument.
|
| Scan protocol |
Slides were scanned using Illumina iScanner
|
| Description |
hiPS-R4
Male, hiPSC, passage 20, derived using lentiviral constructs separately expressing OSKM, grown in feeder-free conditions (vitronectin-coated plates and Essential 8 media)
|
| Data processing |
The data were analyzed with GenomeStudio using Illumina default analysis settings
Quantile normalized and analyzed using Partek's Genomics Suite
|
| |
|
| Submission date |
May 25, 2016 |
| Last update date |
Aug 30, 2017 |
| Contact name |
Mark Ptashne |
| E-mail(s) |
[email protected]
|
| Organization name |
Sloan Kettering Institute
|
| Department |
Molecular Biology Program
|
| Lab |
Mark Ptashne
|
| Street address |
1275 York Ave
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10065 |
| Country |
USA |
| |
|
| Platform ID |
GPL10558 |
| Series (2) |
| GSE81891 |
OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts |
| GSE81900 |
OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts |
|
