|
| Status |
Public on Dec 28, 2008 |
| Title |
N2_d6 #2 (251508710476 Array1_2) |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
N2 day6
|
| Organism |
Mus musculus |
| Characteristics |
embryonic stem cell
Strain: 129/Sv
Gender: male
Age: TBE
Individual Identifier: N2-9 day6 251508710476 Array1_2
|
| Biomaterial provider |
Kazuhiro Aiba,Laboratory of Genetics, NIA/NIH. aibaka@mail.nih.gov, kom@mail.nih.gov
|
| Treatment protocol |
ES cells were plated at 1 x104 cells/cm2 onto gelatin-coated 60 mm-dishes and were cultured as a monolayer in N2B27, which is a 1:1 mixture of DMEM/F12 supplement with modified N2 and Neurobasal medium supplement with B27 without vitamin A (GIBCO) for 6 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted with TRIzol reagent (Invitrogen) according to the manufacturers instructions.
|
| Label |
Cy3
|
| Label protocol |
Total RNA was labeled using Agilents Fluorescent Linear Amplification kit, according to manufacturers instructions (Product Number G2554A or P/N G2556-66002, Version 3.0, June 2002).
|
| |
|
| Channel 2 |
| Source name |
Universal Mouse Reference 129ES Cell
|
| Organism |
Mus musculus |
| Characteristics |
Universale Mouse Reference
|
| Biomaterial provider |
Stratagene
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cultured cells using a Stratagene Absolutely RNA kit. DNase-treated total RNA samples from 11 cultured cell lines derived from embryo, embryo fibroblast, kidney, liver/hepatocyte, lung/alveolar macrophage, B-lymphocyte, T-lymphocyte (thymus), mammary gland, muscle myoblast, skin, and testis were pooled in equal parts. The resulting URM RNA was mixed with 129ES cell RNA (Lif+) at 2:1 ratio and 2.5ug of mixed RNA was used for labeling in each tube
|
| Label |
Cy5
|
| Label protocol |
Total RNA was labeled using Agilents Low RNA Input Fluorescent Linear Amplification kit, according to manufacturers instructions.
|
| |
|
| |
| Hybridization protocol |
Slides were hybridized according to manufacturers protocol (Two-Color Microarray-Based Genr Expression Analysis Protocol, Product # G4140-90050, Version 5.0.1, August 2006)
|
| Scan protocol |
Slides were scanned with an Agilent DNA Microarray Scanner (model G2505-64120) at 100% and 10% PMT in both channels, with a scan resolution of 5um.
|
| Description |
N2 day6
|
| Data processing |
Data are extracted with Agilent Feature Extraction Software.The data were further processed with NIA ANOVA tool utilities.See http://lgsun.grc.nia.nih.gov/ANOVA for details.
|
| |
|
| Submission date |
May 21, 2008 |
| Last update date |
Dec 28, 2008 |
| Contact name |
Minoru S.H. Ko |
| E-mail(s) |
kom@mail.nih.gov
|
| Phone |
410-558-8359
|
| Organization name |
NIH
|
| Department |
National Institute on Aging
|
| Lab |
Lab of Genetics
|
| Street address |
251 Bayview Blvd, Suite 100, 10C
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21224 |
| Country |
USA |
| |
|
| Platform ID |
GPL6867 |
| Series (1) |
| GSE11523 |
Defining Developmental Potency and Cell Lineage Trajectories by Expression Profiling of Differentiating Mouse ES Cells |
|
