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Status |
Public on Jul 31, 2017 |
Title |
somatic cell from aged donor [A-SC-1.4] |
Sample type |
RNA |
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|
Source name |
somatic cell from aged donor
|
Organism |
Mus musculus |
Characteristics |
cell type: somatic cell age: aged
|
Treatment protocol |
Fibroblasts were collected from B6CBAF1 mouse E17.5 embryonic skin and 1.5-year-old tail tip skin. 10E6 fibroblasts were infected with retrovirus generated from pMX-mOCT4, pMX-mSOX2, pMX-mKLF4, and pEYK-mMYC in 6-well dishes with 0.5 ml of each viral supernatant (total 2 ml per well) and spun at 2500 rpm at RT for 90 min (BenchTop Centrifuge, BeckmanCoulter, Allegra-6R). For the generation of A-iPSC-ZSCAN10, the procedure was identical but in addition to the four reprogramming factors, we added a doxycycline-inducible system to overexpress ZSCAN10.”
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Growth protocol |
Mouse ESC and iPSC were cultured in ESC media containing 15% FBS and 1,000 U/ml of LIF (ESGRO® Leukemia Inhibitory Factor [LIF], 1 million units/1 mL). Doxycycline-inducible ZSCAN10 was induced in media supplemented with 2 μg/ml doxycycline (MP Biomedicals, doxycycline hyclate). Human iPSC were cultured in human ESC media (DMEM/F12, 20% knockout serum replacement, 10 ng/ml bFGF, and 0.1 mM -mercaptoethanol) on irradiated mouse embryonic feeders. Human and mouse fibroblasts were cultured in DMEM media with 10% FBS.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was reverse transcribed using the M-MuLV Reverse Transcriptase system (New England Biolabs), and the resulting cDNA was diluted. Synthesized cDNA solution was used for analysis.
|
Label |
cy3
|
Label protocol |
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
|
|
|
Hybridization protocol |
Standard Illumina hybridization protocol
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Scan protocol |
Standard Illumina scanning protocol
|
Data processing |
Data normalization was calculated with the Quantile normalization algorithm implemented in the BioConductor lumi package.
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Submission date |
Jan 10, 2017 |
Last update date |
Jul 31, 2017 |
Contact name |
Hu Li |
E-mail(s) |
li.hu@mayo.edu
|
Organization name |
Mayo Clinic
|
Department |
Molecular Pharmacology & Experimental Therapeutics
|
Lab |
Systems Biology and Pharmacology
|
Street address |
200 First Street, Gonda Building G19-408
|
City |
Rochester |
State/province |
MN |
ZIP/Postal code |
55904 |
Country |
USA |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE85365 |
Recovery of genomic stability by ZSCAN10 in induced pluripotent stem cells from aged donors |
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