|
| Status |
Public on Mar 16, 2015 |
| Title |
ID8_peritoneal cavity selected_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse ovarian cancer cell line_ peritoneal cavity selected
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
gender: female
tissue: ovary
cell line: In vivo-selected ID8 ovarian cancer cells
chip #: 9249051062
position on chip: E
|
| Treatment protocol |
NA
|
| Growth protocol |
Ovarian epithelial papillary serous adenocarcinoma cell line ID8 and in vivo selected ID8-M were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM, GIBCO, Grand Island, NY) supplemented with 4% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, 5 μg/ml insulin, 5 μg/ml transferrin and 5 ng/ml sodium selenite at 37 °C in a humidified 5% CO2 atmosphere.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
miRNeasy Mini Kit (Qiagen) was used to extract Total RNAs according to following instructions: 1. Homogenize cultured cells with 1 ml of “Qiazol” and incubate for 5 min at room temperature; 2. Add 200 ul of chloroform and cap tube securely, vortex vigorously for 15 s, incubate for 2-3 min at room temperature; 3. Centrifuge for 15 min at 14,000 g at 4°C; 4. Transfer the upper aqueous phase to a new collection tube (Avoid transferring any Interphase), add 1.5 volumes of 100% ethanol, and mix thoroughly by shaking; 5. Pipet up to 700 ul sample, including any precipitate, into an RNeasy Mini column in a 2 ml collection tube. Close the lid and centrifuge at ≥ 8000 g for 15s at room temperature; 6. Discard the flow-through; 7. Repeat step 5 using the remainder of the sample; 8. Add 700 ul Buffer RWT to the RNeasy Mini column. Close the lid, and centrifuge for 15 s at ≥ 8000 g. Discard the flow-through; 9. Pipet 500 ul Buffer RPE onto the RNeasy Mini column. Close the lid, and centrifuge for 15 s at ≥8000 g. Discard the flow-through; 10. Add 500 ul Buffer RPE to the RNeasy Mini column. Close the lid, and centrifuge for 2 min at ≥8000 g; 11. Place the RNeasy Mini column into a new 2 ml collection tube, centrifuge at full speed (17,000 g) for 1 min to further dry the membrane. Then open the lid, remove the additional liquid on the walls using pippet; 12. Transfer the RNeasy Mini column to a new 1.5 ml collection tube. Pipet 30 ul RNase-free water directly onto the RNeasy Mini column membrane. Close the lid, and centrifuge for 1 min at ≥8000 g to elute.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
ID8-M2
replicate 2
Total RNA, including microRNA
|
| Data processing |
The data were normalized using quantile normalization with IlluminaGUI in R
|
| |
|
| Submission date |
Mar 16, 2015 |
| Last update date |
Mar 16, 2015 |
| Contact name |
yun lian |
| E-mail(s) |
yun.lian@utsouthwestern.edu
|
| Organization name |
UT Southwestern Medical Center
|
| Street address |
5323 Harry Hines Blvd.
|
| City |
Dallas |
| State/province |
TX |
| ZIP/Postal code |
75390 |
| Country |
USA |
| |
|
| Platform ID |
GPL6887 |
| Series (1) |
| GSE66934 |
In vivo-selected ID8 ovarian cancer cells display accelerated peritoneal metastasis accompanied with increased macrophage recruitment and M2 polarization |
|
