|
| Status |
Public on Apr 14, 2020 |
| Title |
Rptor WT_5 |
| Sample type |
RNA |
| |
|
| Source name |
wild-type epidermis
|
| Organism |
Mus musculus |
| Characteristics |
genotype/variation: wild-type
age: E18.5
tissue: epidermis
|
| Growth protocol |
Mouse epidermal tissue dissected from E18.5 mouse embryos
|
| Extracted molecule |
total RNA |
| Extraction protocol |
E18.5 embryonic littermates were manually classified as wild-type or Rptor cKO and were additionally validated by genotyping. Mouse epidermis was separated from the dermis following incubation of E18.5 embryonic skin with 3.8% Ammonium thiocynate for 10 minutes at room temperature. The epidermal sheet was homogenized in TRIzol and total RNA prepared using the TRIzol RNA extraction and RNeasy purification protocol.
|
| Label |
biotin
|
| Label protocol |
The Ambion Expression WT kit was used as described in the manufacturer's manual.
|
| |
|
| Hybridization protocol |
17hrs at 45° C with rotation (60rpm) as described by Affymetrix in their GeneChip Expression Analysis Technical Manual.
|
| Scan protocol |
Biotin labeling was performed using the GeneChip WT Pico Kit (Life Technologies, Grand Island, NY) according to the manufacturer's protocol, followed by the GeneChip WT Terminal Labeling and Controls Kit (Affymetrix, Santa Clara, CA).
|
| Description |
RAPTOR WT#5
|
| Data processing |
Affymetrix CEL files were extracted and their data normalized with the Partek GS 6.6 platform, with junction probes, annotated as JUC~ in the meta-probeset file, excluded. Affymetrix's na35 probeset set was used with RMA normalization to create quantile-normalized log2 transcript signal values, which were used in subsequent ANOVA analyses.
probe group file: MTA-1_0.r3.pgf
meta-probeset file: MTA-1_0.na35.mm10.probeset.csv
|
| |
|
| Submission date |
Jan 07, 2019 |
| Last update date |
Apr 14, 2020 |
| Contact name |
KAUSHAL VIJAY ASRANI |
| E-mail(s) |
[email protected]
|
| Phone |
4109268061
|
| Organization name |
JOHNS HOPKINS MEDICINE
|
| Street address |
1550 ORLEANS
|
| City |
BALTIMORE |
| State/province |
MD |
| ZIP/Postal code |
21231 |
| Country |
USA |
| |
|
| Platform ID |
GPL20775 |
| Series (1) |
| GSE124754 |
mTORC1 feedback to AKT modulates MiT/TFE-driven lysosomal biogenesis and EGFR degradation |
|
