|
Status |
Public on Aug 29, 2007 |
Title |
ORMES22_ReplicateA |
Sample type |
RNA |
|
|
Source name |
Rhesus monkey IVF-derived embryonic stem cell line ORMES22 biological replicate A
|
Organism |
Macaca mulatta |
Characteristics |
Undifferentiated rhesus monkey IVF-derived Oregon Rhesus Macaque Embryonic Stem (ORMES) cell line 22 biological replicate A.
|
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)
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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).
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|
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Submission date |
May 04, 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 |
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