|
| Status |
Public on Jun 25, 2007 |
| Title |
Meis_2 |
| Sample type |
RNA |
| |
|
| Source name |
nephrogenic interstitium
|
| Organism |
Mus musculus |
| Characteristics |
gene reported: Tg(Meis1-EGFP)FO156Gsat / MGI:3843887
strain: CD-1
sex: Unknown
developmental stage: E15.5
theiler stage: 23
somite count: NA
developmental landmark: NA
|
| Treatment protocol |
pool size: approx 20 E15.5 kidneys
Pooled sample: Yes
Dissection Method: Fluorescent Activated Cell Sorting (FACS)
Meis transgenic mice are time-mated. Pregnant Meis 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 dissociated cells from the kidneys are pelleted at 5000 rpm, 4 ºC for 5 minutes. The media is aspirated and the cell pellet is resuspended in 200µl of ice-cold 2%FBS/PBS. Filter cells through 70 micron mesh filter and the GFP positive cells are further isolated using FACS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Potter protocol: 'RNA purification'
|
| Label |
Biotin
|
| Label protocol |
Biotin-X-X-NHS (EPICENTRE Biotechnologies)
|
| |
|
| Hybridization protocol |
Affymetrix standard protocol
Amount labeled target hybridization to array: 5ug
|
| Scan protocol |
Affymetrix standard protocol
|
| Description |
>> Amplification protocol <<
Target Amplified manufacturer/kit: TargetAmp 2-Round Aminoallyl-aRNA Amplification Kit 1.0 (EPICENTRE Biotechnologies)
Target Amplified protocol: Potter protocols
Rounds of amplification: 2
URL: http://gudmap.hgu.mrc.ac.uk/gudmap/pages/mic_submission.html?id=GUDMAP:8101
|
| Data processing |
Analysis method: Affymetrix GCOS and Gene Spring and Avadis programs GCOS Tgt value: 1500
|
| |
|
| Submission date |
Jun 22, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
GUDMAP Developers |
| E-mail(s) |
gudmap-db@gudmap.org
|
| 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 |
GPL1261 |
| Series (1) |
| GSE8232 |
Gene expression profiles of E15.5 peripheral blastema isolated from Meis1-EGFP transgenic mice using FACS. (GUDMAP Series ID: 17) |
|
| Relations |
| Reanalyzed by |
GSE119085 |
