|
| Status |
Public on Feb 01, 2012 |
| Title |
12FlkM2 |
| Sample type |
RNA |
| |
|
| Source name |
Gonad, 12.5 dpc male endothelial cells
|
| Organism |
Mus musculus |
| Characteristics |
gene reported: Tg(Kdr-mCherry)1Medi/MGI:4417934
strain: CD-1
Sex: male
developmental stage: 12.5 dpc
theiler stage: TS20
|
| Treatment protocol |
Pooled sample: Yes
Pool size: one or more litters
Dissection method: Dissected out the gonad, trypsinized and FACS sorted. Capel protocol: FACS Protocol
Experimental design: Female mice were time-mated and the embryos were collected. The gonads were removed and the males and females were separately pooled. The gonads were incubated in Trypsin EDTA for 5-10 minutes. The trypsin was removed, PBS (often with DNase) was added, and the gonadal cells were dissociated by pipetting and pulling through a syringe. The samples were passed through a strainer and then FACS sorted. Capel protocol: FACS protocol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Capel protocol: RNA extraction and sample preparation for Affymetrix Gene 1.0 ST arrays
|
| Label |
Biotin
|
| Label protocol |
Nugen Encore Biotin Module (4200)
|
| |
|
| Hybridization protocol |
Affymetrix standard protocol
The arrays were washed and stained using the Affymetrix Fluidics Station 450. Fluidics Protocol FS450-0007.
Amount labeled target hybridization to array: Approximately 2.5 µg into the hybridization cocktail, approximately 2.1 µg were hybridized to the array.
|
| Scan protocol |
Probe arrays were scanned using an Affymetrix GeneChip Scanner 3000 7G and Affymetrix GeneChip Command Console (AGCC) Software version 3.1.1.
|
| Description |
Amplification protocol
Target Amplified manufacturer/kit: Nugen WT-Ovation Pico RNA Amplification System (3300) and WT-Ovation Exon Module (2000)
Target Amplified protocol: Capel protocol: RNA extraction and sample preparation for Affymetrix Gene 1.0 ST arrays
Rounds of amplification: 1
URL: http://www.gudmap.org/gudmap/pages/mic_submission.html?id=GUDMAP:14006
|
| Data processing |
Analysis method: Partek
GCOS Tgt value: 500
|
| |
|
| Submission date |
Mar 07, 2011 |
| Last update date |
Feb 01, 2012 |
| 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) |
| GSE27715 |
Gene expression profiles of germ cells, supporting cells, interstitial cells (including steroidogenic precursors), and endothelial cells in the developing testis and ovary at 11.5, 12.5, and 13.5 dpc. (GUDMAP Series ID: 43) |
|
