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Status |
Public on Jul 16, 2010 |
Title |
CRYM_P1 |
Sample type |
RNA |
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|
Source name |
cap mesenchyme
|
Organism |
Mus musculus |
Characteristics |
gene reported: Tg(Crym-EGFP)82Gsat / MMRRC:012003-UCD strain: CD-1 sex: unknown developmental stage: P1 theiler stage: TS27 somite count: NA developmental landmark: NA
|
Treatment protocol |
pool size: approx 4 P1 kidneys Pooled sample: Yes Dissection Method: Trypsinization and fluorescent activated cell sorting (FACS). Crym transgenic mice were time-mated. Pregnant Crym transgenic mice were euthanized by standard carbon dioxide asphyxiation. Neonatal pups were fostered to new CD-1 females. At the appropriate time, pups were killed by decapitation with a scalpel and the kidneys were isolated and placed in ice-cold PBS. The kidneys were minced with a razor blade and incubated in the presence of 300µl of 0.25% trypsin-EDTA for 5 minutes at 37ºC. After incubation, the kidneys were further dissociated by titurating in the presence of 600µl of ice-cold 10%FBS/PBS, and then pelleted at 5000 rpm, 4ºC for 5 minutes. The cells were resuspended in 200µl of ice-cold 2%FBS/PBS, filtered using a 70 micron mesh and isolated using 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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|
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Hybridization protocol |
Affymetrix standard protocol Amount labeled target hybridization to array: 2.5 µg The arrays were washed and and stained using a Fluidics Station 450 (Affymetrix) utilizing the fluidics protocol FS450-0007.
|
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:12315
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Data processing |
Analysis method: Affymetrix Expression Console and GeneSpring 10.
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|
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Submission date |
Jul 16, 2009 |
Last update date |
Dec 31, 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 (2) |
GSE17139 |
Gene expression profiles of cap mesenchyme and renal vesicle isolated between P0-P4 from Crym-EGFP neonatal transgenic mice using FACS. (GUDMAP Series ID: 28) |
GSE17218 |
Encyclopedia of the expression levels of all genes in multiple components of the developing kidney |
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