|
| Status |
Public on Nov 21, 2011 |
| Title |
FACS-sorted embryonic trachea, midline cells, ttk mutant 1: ttk2D50 btl-enh-RFPmoe |
| Sample type |
RNA |
| |
|
| Source name |
btl-enhancer embryonic expression domain, FACS-sorted
|
| Organism |
Drosophila melanogaster |
| Characteristics |
strain: ttk2D50 btl-enh-RFPmoe
tissue: Tracheal system, midline
|
| Treatment protocol |
For each condition, 100.000 btl RFP moe positive cells sorted out by fluorescence activated cell sorting were used to extract their RNA.
|
| Growth protocol |
Embryos btl RFP moe ttk/twi GFP and btl RFP moe/twi GFP were grown at 25C for 7:40 hrs and then age for 5:20 hr more. The overexpression condiiton stock UAS ttk btl RFP moe x Btl Gal4 UAS tau GFP and its control stock UAS ttk RPF moe were grown at 29C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with a combination of Trizol (for the whole embryo condition) and RNeasy minikit (Qiagen)(for the btl RFP positive cells)
|
| Label |
biotin
|
| Label protocol |
25 ng total RNA per sample was processed using isothermal amplification SPIA Biotin System (NuGEN Technologies, Inc.)
|
| |
|
| Hybridization protocol |
2.2 µg of cDNA was hybridized per microarray for 16 hours of hybridization at 45C, washing and staining of microarrays was performed using a Fluidics Station 450 (Affymetrix, Santa Clara, CA)
|
| Scan protocol |
GeneChips were scanned in a GeneChip Scanner 3000 (Affymetrix, Santa Clara, CA)
|
| Data processing |
Data from ttk mutants and ttk overexpression were pre-processed separately. Data was processed using ENSEMBL gene-specific Custom CDF V12 (brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/genomic_curated_CDF.asp), quantile normalized and RMA summarized using Bioconductor.
|
| |
|
| Submission date |
Jun 27, 2011 |
| Last update date |
Nov 21, 2011 |
| Contact name |
Boris Adryan |
| Organization name |
University of Cambridge
|
| Department |
Cambridge Systems Biology Centre & Department of Genetics
|
| Lab |
Adryan Lab
|
| Street address |
Tennis Court Road
|
| City |
Cambridge |
| ZIP/Postal code |
CB2 1QR |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL13770 |
| Series (1) |
| GSE30239 |
Mis-expression of Tramtrack in Drosophila embryos |
|
