|
| Status |
Public on Dec 09, 2015 |
| Title |
201B7_p45 |
| Sample type |
RNA |
| |
|
| Source name |
201B7, passage 45, untreated
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: human HDF-derived iPS cell
cell line: 201B7
passage number: 45
lectin treatment: none
treatment period: 0
|
| Growth protocol |
Human iPSCs and ESCs were maintained in an undifferentiated state in the presence of bFGF. The cultures were incubated at 37 ºC in a humidified incubator with 3 or 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using ISOGEN (Nippon Gene) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 150 ng of total RNA using Low Input Quick Amp Labeling Kit (Agilent) according to the manufacturer's instructions, followed by RNeasy column purification (QIAGEN). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
0.6 ug of Cy3-labelled cRNA (specific activity >6.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 25 ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x HiRPM Hybridization Buffer was added to the fragmentation mixture and hybridized to Agilent SurePrint G3 Human GE 8x60K Microarray Kit (G4851A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x60k array slides.
|
| Description |
Gene expression in human iPS cell
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software GX12.0 (Agilent) using default parameters.
|
| |
|
| Submission date |
Dec 18, 2012 |
| Last update date |
Dec 09, 2015 |
| Contact name |
Yuzuru Ito |
| Organization name |
National Institute of Advanced Industrial Science and Technology (AIST)
|
| Department |
Biotechnology Research Institute for Drug Discovery
|
| Street address |
Central 6, Higashi 1-1-1
|
| City |
Tsukuba |
| State/province |
Ibaraki |
| ZIP/Postal code |
3058566 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13607 |
| Series (1) |
| GSE42976 |
Effect of rBC2LCN on human embryonic stem cells |
|
