|
| Status |
Public on Dec 05, 2012 |
| Title |
ES Activin (-) 2 |
| Sample type |
RNA |
| |
|
| Source name |
EB3_control
|
| Organism |
Mus musculus |
| Characteristics |
strain background: 129/Ola
cell line: EB3
cell type: embryonic stem cells
molecule subtype: miRNA
|
| Treatment protocol |
Mouse ES cells were treated with Activin 10ng/ml for 18 days.
|
| Growth protocol |
Mouse ES cells were differentiated through 3 days formation of embryoid bodies with Activin 10ng/ml. After embryoid body formation, embryoid bodies were transfeterd onto cell culture dishes coated with gelatin. Cells were subsequently cultured with Activin 10ng/ml for additionally 15 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.And RNA was prepared using the , miRNAeasy mini (QIAGEN, Valencia, CA) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
pCp Cyanine-3 (Cy3) labeled miRNA was prepared from 100ng total RNA using the miRNA Complete Labeling Kit(Agilent) according to the manufacturer's instructions.After labering reaction,completely dray the samples.Use a vacuum concentrator at the midium-high heat setting.. Resuspend the dried sample in 18ul of nuclease-free water.
|
| |
|
| Hybridization protocol |
18ul of Cy3-labelled miRNA was fragmented following the manufacturers instructions. On completion of the fragmentation reaction, 4.5ul of 10x GE Block and 22.5ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Mouse miRNA Microarray Kit (V2), 8x15K (Based on Sanger miRBase release 12.0)ver2(G4471A) for 20 hours at 55℃,20RPM in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent) and 5 minute at 37℃ GE Wash buffer 2 (Agilent), then dried immediately by air spray
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565CA) using one color scan setting for 8x15k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to XDR Hi 100% and XDR Lo 5% ).
|
| Description |
SAMPLE 2
|
| Data processing |
The scanned images were analyzed with Feature Extraction software Ver 10.1.1(Agilent).
|
| |
|
| Submission date |
Dec 04, 2012 |
| Last update date |
Dec 05, 2012 |
| Contact name |
Ryuji Morizane |
| E-mail(s) |
[email protected]
|
| Organization name |
Keio University School of Medicine
|
| Department |
Internal Medicine
|
| Lab |
Renal
|
| Street address |
35 shinanomachi, shinjukuku
|
| City |
Tokyo |
| ZIP/Postal code |
1608582 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10384 |
| Series (2) |
| GSE42717 |
Mouse ES cell differentiated with Activin |
| GSE42719 |
miR-34c attenuates epithelial-mesenchymal transition and renal fibrosis of kidneys with ureteral obstruction. |
|
