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Status |
Public on Nov 21, 2011 |
Title |
Whole-embryo, ttk mutant 1: ttk2D50 btl-enh-RFPmoe |
Sample type |
RNA |
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Source name |
whole embryo
|
Organism |
Drosophila melanogaster |
Characteristics |
strain: ttk2D50 btl-enh-RFPmoe tissue: whole embryo
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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.
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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.)
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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)
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Scan protocol |
GeneChips were scanned in a GeneChip Scanner 3000 (Affymetrix, Santa Clara, CA)
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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.
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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 |
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|
Platform ID |
GPL13770 |
Series (1) |
GSE30239 |
Mis-expression of Tramtrack in Drosophila embryos |
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