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Status |
Public on Mar 23, 2010 |
Title |
Tie2_E15.5_1 |
Sample type |
RNA |
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Source name |
small blood vessels
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Organism |
Mus musculus |
Characteristics |
gene reported: Tg(TIE2GFP)287Sato / MGI:3056937 strain: CD-1 developmental stage: 15.5 dpc theiler stage: TS23
|
Treatment protocol |
pool size: 4-5 Embryonic Kidneys Pooled sample: Yes Dissection Method: trypsinization & fluorescent activated cell sorting Tie2-GFP transgenic mice are time-mated. Pregnant Tie2-GFP transgenic mice are euthanized by standard carbon dioxide asphyxiation. All fetuses are killed by decapitation with a scalpel. Fetal kidneys are dissected from fetuses and placed in ice-cold PBS. Embryonic kidneys are incubated in the presence of 300µl of trypsin for 5 minutes at 37 ºC. Kidneys are then dissociated by titurating in the presence of 600µl of ice-cold 10%FBS/PBS. The kidneys are pelleted at 5000 rpm, 4 ºC for 5 minutes. The media is aspirates and the cell pellet is resuspended in 200µl of ice-cold 2%FBS/PBS. Filter cells through 70 micron mesh filter and proceed with FACS.
|
Extracted molecule |
total RNA |
Extraction protocol |
Potter protocol: 'RNA purification'
|
Label |
Biotin
|
Label protocol |
The FL-Ovation cDNA Biotin Module V2 (NuGEN) is used to chemically modify fragments and label target sense cDNA.
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Hybridization protocol |
Affymetrix standard protocol Amount labeled target hybridization to array: 2.5µg The arrays were washed and and stained using a Fluidics Station 450 (Affymetrix) utilizing the fluidics protocol FS450-0007.
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Scan protocol |
Probe arrays were scanned using an Affymetrix GeneChip Scanner 3000 7G and Affymetrix scanning software Genechip Operating Software Version 1.4.
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Description |
>> Amplification protocol << Target Amplified manufacturer/kit: WT-Ovation Pico RNA Amplification System (NuGEN) Target Amplified protocol: Potter protocols Rounds of amplification: 1 URL: http://www.gudmap.org/gudmap/pages/mic_submission.html?id=GUDMAP:13556
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Data processing |
Analysis method: Affymetrix Expression Console and GeneSpring 10.
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Submission date |
Mar 22, 2010 |
Last update date |
Dec 30, 2010 |
Contact name |
GUDMAP Developers |
E-mail(s) |
[email protected]
|
Phone |
+44 131 651 8500
|
Organization name |
IGMM MRC Human Genetics Unit
|
Lab |
GUDMAP Database Group
|
Street address |
Crewe Road
|
City |
Edinburgh |
ZIP/Postal code |
EH4 2XU |
Country |
United Kingdom |
|
|
Platform ID |
GPL6246 |
Series (1) |
GSE20991 |
Gene expression profiles of E15.5 endothelial cells isolated from TIE2-GFP transgenic mice using FACS. (GUDMAP Series ID:38) |
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