|
| Status |
Public on Dec 14, 2016 |
| Title |
iPSC-CM_donorE_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
iPSC-CM_donorE
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: iPSC-CM
donor: E
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was purified with an RNeasy Mini Kit (Qiagen) following the manufacturer's protocol.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.1 ug RNA using the One-Color Low RNA Input Quick Amplification Labeling kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy 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 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent SurePrint G3 Human GE v2 8x60K Microarray (G039494) 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 (G2505B) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
E5CM-2
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.1.1 (Agilent) using default parameters (protocol GE1_107_Sep09 and Grid: 039494_D_F_20140813) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Dec 14, 2016 |
| Last update date |
Dec 14, 2016 |
| Contact name |
Tadahiro Shinozawa |
| E-mail(s) |
[email protected]
|
| Organization name |
Takeda pharmaceuticals
|
| Street address |
Muraokahigashi 2chome
|
| City |
Fujisawa |
| ZIP/Postal code |
251-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE92391 |
Global gene expression of hiPS cell derived cardiomyocytes from healthy volunteers |
|
