|
| Status |
Public on Aug 19, 2015 |
| Title |
minKGFP_high_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
E10.5 minKGFP high conduction system cells
|
| Organism |
Mus musculus |
| Characteristics |
transgenic line: minKGFP
Stage: E10.5
|
| Growth protocol |
Hearts from E10.5 minKGFP transgenic embryos were isolated and digested to single cells with pancreatin and collagenase. Cells were FACS sorted into minKGFP high and minKGFP low groups on the basis of fluoresence intensity.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were sorted directly into Trizol LS (Invitrogen), and RNA was extracted according to the manufacturer's instructions.
|
| Label |
AlexaFluor-3
|
| Label protocol |
5 ng of total RNA from each sample was amplified with the NuGen WT Ovation Pico kit according to the manufacturer's instructions. 2 ug of amplified product was subsequently labeled with AlexaFluor-3 using the BioPrime Total Labeling Kit (Invitrogen) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
3.26 ug of AlexaFluor-3 labeled sample in a volume of 24 uL was added to 6 uL of 10x blocking agent (Agilent) and 30 uL of 2x hybridization buffer (Agilent) for a final volume of 60 uL. 55 uL was added to a Tecan HS 4800 Pro chamber and hybridized to Agilent G4122F mouse whole genome microarrays using a standard 17 hour 65C gene expression hybridization.
|
| Scan protocol |
Slides were scanned after washing on the Agilent Technologies Scanner G2505C using one color settings for 4 x 44k arrays at a resolution of 5 microns per pixel.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.5.1.1 (Agilent) using default parameters (protocol GE1_105_Dec08 and Grid 014868_D_F_20090416).
|
| |
|
| Submission date |
Feb 28, 2012 |
| Last update date |
Aug 19, 2015 |
| Contact name |
David Arnolds |
| E-mail(s) |
dea@uchicago.edu
|
| Organization name |
University of Chicago
|
| Department |
Pediatrics
|
| Lab |
Moskowitz
|
| Street address |
920 E 57th St
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60637 |
| Country |
USA |
| |
|
| Platform ID |
GPL4134 |
| Series (1) |
| GSE36136 |
Expression profiling of the E10.5 cardiac conduction system using MinKGFP BAC transgenic mice |
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