|
| Status |
Public on Jul 29, 2014 |
| Title |
ShhCre control lung -biological rep 3 |
| Sample type |
RNA |
| |
|
| Source name |
E12.5 Lung, Shhcre control
|
| Organism |
Mus musculus |
| Characteristics |
genotype: Shh cre/+
tissue: embryonic lung
age: E12.5
|
| Treatment protocol |
Lung buds were disected from E12.5 embryos with appropriate genotypes and stored in Trizol for future RNA extraction
|
| Growth protocol |
Timely matings were set up to obtain E12.5 embryos
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using Rneasy mini kit (Qiagene)
|
| Label |
Biotin
|
| Label protocol |
100ng of total RNA was converted to first-strand cDNA using reverse transcriptase primed by a poly(T) oligomer that incorporated the T7 RNA polymerase promoter. Second-strand cDNA synthesis was followed by in vitro transcription (Affymetrix One-Cycle Target Labeling Kit) for linear amplification and biotinylation of each transcript.
|
| |
|
| Hybridization protocol |
cDNAs were added to Affymetrix hybridization cocktails, heated at 99ºC for 2 min and hybridized for 16 h at 45ºC to GeneChips (Affymetrix Inc., Santa Clara CA). The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin. Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain.
|
| Scan protocol |
A confocal scanner was used to collect fluorescence signal after excitation at 570 nm.
|
| Data processing |
The data were analyzed using scripts written in the R languge using the Bioconductor package Oligo using the default settings for RMA.
|
| |
|
| Submission date |
Jul 30, 2013 |
| Last update date |
Jul 29, 2014 |
| Contact name |
Rachel S Kadzik |
| E-mail(s) |
rkadzik@mail.med.upenn.edu
|
| Organization name |
University of Pennsylvania
|
| Department |
Molecular Cardiology
|
| Lab |
Edward E Morrisey
|
| Street address |
Translational Research Center, 11th Flr.; Room 179, 3400 Civic Center Boulevard, Bldg 421
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104-5159 |
| Country |
USA |
| |
|
| Platform ID |
GPL16570 |
| Series (1) |
| GSE49370 |
Role of Fzd2 in lung epithelium development |
|
