|
| Status |
Public on Feb 10, 2012 |
| Title |
CBFb_KD_1 |
| Sample type |
RNA |
| |
|
| Source name |
CBFb_KD
|
| Organism |
Mus musculus |
| Characteristics |
cell line: L8057
cell type: megakaryocytic cells
genotype/variation: shRNA-mediated CBFb knockdown
|
| Treatment protocol |
48 hrs after lenti-viral infection, 2 microgram/microliter puromycin was added to culture medium for selecting infected cells, and cells were harvested for RNA 72 hrs after puromycin selection. 50 nM TPA was added at the same time with puromycin to induced maturation of the cells.
|
| Growth protocol |
Cells were grown in 38.5% IMDM + 38.5% RPMI + 20% FCS + %2 Pen/strep + 1% L-Glutamine,
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy Plus Mini Kit (Qiagen, catalogue no. 74134).
|
| Label |
biotin
|
| Label protocol |
5 microgram RNA is converted into cDNA using a T7 promoter-tailed oligo-dT primer in the synthesis of the first cDNA strand; second strand cDNA synthesis is then carried out. The double-stranded cDNA is used as the template in an in vitro transcription (IVT) reaction catalyzed by T7 polymerase and containing biotinylated CTP and UTP in addition to the four unmodified ribonucleoside triphosphates.
|
| |
|
| Hybridization protocol |
hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 Arrays (MOE430v2) following standard techniques.
|
| Scan protocol |
Scanned with GeneChIP scanner 300 7G using the standard protocol.
|
| Description |
maturation was induced by adding 50 nM (final concentration) 12-O- tetradecanoylphorbol-13-acetate (TPA) to the medium for 3 days
|
| Data processing |
Expression data were analyzed using R and Bioconductor (http://www.bioconductor.org/).
|
| |
|
| Submission date |
Nov 14, 2011 |
| Last update date |
Feb 10, 2012 |
| Contact name |
Tali Mazor |
| Organization name |
MIT
|
| Street address |
77 Massachusetts Ave
|
| City |
Cambridge |
| State/province |
MA |
| ZIP/Postal code |
02139 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (2) |
| GSE33659 |
Direct Recruitment of Polycomb Repressive Complex 1 (PRC1) to Chromatin by Core Binding Transcription Factors (microarray) |
| GSE33660 |
Direct Recruitment of Polycomb Repressive Complex 1 (PRC1) to Chromatin by Core Binding Transcription Factors |
|
