|
| Status |
Public on Aug 29, 2007 |
| Title |
ORMES22_ReplicateB |
| Sample type |
RNA |
| |
|
| Source name |
Rhesus monkey IVF-derived embryonic stem cell line ORMES22 biological replicate B
|
| Organism |
Macaca mulatta |
| Characteristics |
Undifferentiated rhesus monkey IVF-derived Oregon Rhesus Macaque Embryonic Stem (ORMES) cell line 22 biological replicate B.
|
| Biomaterial provider |
James A. Byrne, PhD
|
| Treatment protocol |
Embryonic stem cells with good morphology were manually removed from culture for total RNA purification.
|
| Growth protocol |
Embryonic stem cells were grown at 37°C in 5% CO2 atmosphere on mitotically inactivated mouse embryonic fibroblasts (MEF) feeder cells in Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) (Invitrogen) supplemented with 15% foetal bovine serum (FBS) (Hyclone), 0.1 mM ß-mercaptoethanol (Sigma), 1% nonessential amino acids (Invitrogen), 4ng/ml FGF2, 2 mM L-glutamine (Invitrogen) and a pH of 7.2 (altered with 4N NaOH). ES cell colonies were manually passaged every 4-5 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cell colonies selected for the appropriate ESC morphology (flat monolayer colony with distinctive cobbled stem cell morphology and a high nucleo-cytoplasmic ratio) using the Invitrogen TRIZOL reagent, followed by further purification with the Qiagen RNeasy MinElute Cleanup Kit. The RNA samples were quantified using the NanoDrop ND-1000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE) and the quality of the RNA was assessed using Lab-on-a-Chip RNA Pico Chips and the 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).
|
| Label |
Biotin
|
| Label protocol |
Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual, rev.3. 2001)
|
| |
|
| Hybridization protocol |
Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual, rev.3. 2001)
|
| Scan protocol |
Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual, rev.3. 2001).Scanning Hardware – HP GeneArray Scanner
|
| Description |
Probe level measurements produced by Affymetrix Microarray Suite 5.0. Analysis performed at level of raw signal. (See Affymetrix Microarray Suite 5.0 reference on web site for description).
|
| Data processing |
MAS 5.0; Data was normalized with Standard Affymetrix Protocols (Affymetrix GeneChip Expression Analysis Technical Manual, rev.3. 2001). Two .CHP files are provided as supplementary files: one normalized (_GS.CHP) and one not (.CHP).
|
| |
|
| Submission date |
May 05, 2007 |
| Last update date |
May 07, 2007 |
| Contact name |
James Anthony Byrne |
| E-mail(s) |
byrnej@ohsu.edu
|
| Phone |
5036905335
|
| Organization name |
ONPRC
|
| Department |
Reproductive Sciences
|
| Lab |
Wolf lab
|
| Street address |
505 NW 185th Ave
|
| City |
Beaverton |
| State/province |
OR |
| ZIP/Postal code |
97006 |
| Country |
USA |
| |
|
| Platform ID |
GPL3535 |
| Series (1) |
| GSE7748 |
Transcriptional profiling of rhesus monkey nuclear transfer embryonic stem cells |
|
