|
| Status |
Public on Aug 22, 2017 |
| Title |
Men1-/- MEFs-2 |
| Sample type |
RNA |
| |
|
| Source name |
Men1 knockout Mouse embryonic fibroblasts complemented with vector
|
| Organism |
Mus musculus |
| Characteristics |
strain: mixed FVB;129Sv background
genotype: Men1 knockout
tissue: Mouse embryonic fibroblasts
|
| Treatment protocol |
Seeded cells into 10 cm2 plate at 70% density. After 24 hours incubation at 37C, detached the cells with 0.25% Tripsin and harvested the cells.
|
| Growth protocol |
Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS and 1% Pen/Strep at 37C
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Total RNA was converted to first-strand cDNA using reverse transcriptase primed by poly(T) and random oligomers that incorporated the T7 promoter sequence. Second-strand cDNA synthesis was followed by in vitro transcription with T7 RNA polymerase for linear amplification of each transcript, and the resulting cRNA was converted to cDNA, fragmented, assessed by Bioanalyzer, and biotinylated by terminal transferase end labeling.
|
| |
|
| Hybridization protocol |
15 ug labeled cDNA were added to Affymetrix hybridization cocktails, heated at 99ºC for 5 min and hybridized for 16 h at 45ºC to Affymetrix Murine Genome U74Av2 GeneChips using the GeneChip Hybridization oven 640.
|
| Scan protocol |
The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin. Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain. A GeneChip 3000 7G scanner was used to collect fluorescence signal.
|
| Description |
Gene expression data from Men1 knockout Mouse embryonic fibroblasts complemented with vector
|
| Data processing |
Affymetrix Command Console and Expression Console were used to quantitate expression levels for targeted genes using MAS 5.0
|
| |
|
| Submission date |
Oct 07, 2016 |
| Last update date |
Aug 22, 2017 |
| Contact name |
Xianxin Hua |
| E-mail(s) |
huax@mail.med.upenn.edu
|
| Organization name |
University of Pennsylvania Perelman School of Medicine
|
| Department |
Department of Cancer Biology
|
| Lab |
Hua Lab
|
| Street address |
421 Curie BLVD
|
| City |
philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL81 |
| Series (1) |
| GSE87733 |
Expression data from Men1 knockout MEFs |
|
