|
| Status |
Public on May 21, 2015 |
| Title |
Meg-3_3 |
| Sample type |
RNA |
| |
|
| Source name |
MIN6-4N_Meg3_overexpression
|
| Organism |
Mus musculus |
| Characteristics |
cell line: MIN6-4N
cell type: pancreatic beta cell
|
| Treatment protocol |
MIN6-4N cells were transfected with Vector or Meg3 plasmid using the nucleofection method with Solution -T (Amaxa/Lonza) and selected for 5 weeks in 750 µg/ml G418. Stable clones were picked by using cloning cylinders and expanded, and tested for Meg3 expression by RT-PCR.
|
| Growth protocol |
MIN6-4N stable cell lines cells were cultured in DMEM low glucose medium supplemented with 15% FBS, 1X antibiotic and antimycotic, and 500 µg/ml G418 at 37˚C and 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the RNAeasy kit according to the manufacturer's instructions (Qiagen) from 3 independent cultures of a vector transfected clone (Vec-3), and a Meg3 transfected clone (Meg-5).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Mouse Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
gene expression upon stable mouse Meg3 expression
The sorted cell subpopulations
|
| Data processing |
RMA by software Partek
|
| |
|
| Submission date |
May 15, 2014 |
| Last update date |
May 21, 2015 |
| Contact name |
WeiPing Chen |
| E-mail(s) |
[email protected]
|
| Phone |
301-496-0175
|
| Organization name |
NIDDK/NIH
|
| Department |
GCL
|
| Lab |
Genomics Core Lab
|
| Street address |
Bldg 8, Room 1A11, NIDDK/NIH
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL10740 |
| Series (1) |
| GSE57716 |
Meg3 regulated genes in pancreatic beta cells |
|
