|
| Status |
Public on Oct 28, 2009 |
| Title |
wt lenses at E13.5, biological rep4 |
| Sample type |
RNA |
| |
|
| Source name |
lenses from the wild type embryo at E13.5
|
| Organism |
Mus musculus |
| Characteristics |
tissue: embryonic lenses at E13.5
genotype: Le-Cre-/- ;DicerloxP/loxP
age: mouse embryo at E13.5
|
| Growth protocol |
To generate targeted deletion of Dicer gene, the Le-Cre mice in which the Pax6 P0 promoter drives the expression of Cre recombinase in the surface ectoderm-derived corneal and lens epithelium from E9.0 onward were mated to the mice that had LoxP sites inserted in the Dicer gene (Murchison et al., 2005). The heterozygous progeny were crossed to obtain Le-Cre+/- ;DicerloxP/loxP (DicerCN) and Le-Cre-/-;DicerloxP/loxP (control) offsprings.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Intact lenses were dissected from five control and five DicerCN mice at E13.5 and immediately snap-frozen in liquid nitrogen. The pair of lenses from each mouse was combined for isolation of total RNAs so that five independent samples for each group (WT versus DICER lenses) were submitted for gene expression analysis.
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated sense strand cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA (GeneChip® Whole Transcript (WT) Sense Target Labeling Assay)
|
| |
|
| Hybridization protocol |
5.5 ug of sense strand cDNA were hybridized for 17+ hr at 45C on GeneChip Mouse Gene ST array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneArray Scanner 3000 7G
|
| Description |
Gene expression data from wild type E13.5 embryos
|
| Data processing |
The data were analyzed with GeneChip Operating System (GCOS) using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
May 22, 2009 |
| Last update date |
May 22, 2009 |
| Contact name |
Yan Li |
| E-mail(s) |
liyan2@mail.nih.gov
|
| Phone |
3014962764
|
| Organization name |
NIH\NEI
|
| Lab |
LMDB
|
| Street address |
5625 Fishers Lane, Room 1S-02
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE16209 |
Comparison of lens gene expression between the control and DicerCN mouse embryos at E13.5 |
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