|
| Status |
Public on Aug 07, 2013 |
| Title |
R175Q 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
R175Q 2
|
| Organism |
Mus musculus |
| Characteristics |
treatment: ShcA R175Q expression
cell line: NMuMG-NT2197
|
| Growth protocol |
NMuMG-NT2197 cells were generated, cultured, and transfected with a pMSCV/hygro expression vector (Clontech) expressing a wildtype ShcA cDNA or a ShcA mutant harbouring an arginine to glutamine substitution at amino acid 175 (R175Q)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA (1-2.5ng) from microdissected material was subjected to two rounds of linear amplification using a RiboAmp HSPlus Amplification Kit (Cat. #: KIT0525, Applied Biosystems) following the manufacturer’s protocol.
|
| Label |
Cy3
|
| Label protocol |
The profiles of resulting amplified RNA (aRNA) were assessed using a 2100 Bioanalyzer (Agilent Technologies). The aRNA samples (5μg) were conjugated to Cy3 dye using an Arcturus Turbo Labeling Kit (Cat. #: KIT0609, Applied Biosystems). Universal human reference RNA (Stratagene) was amplified using the same procedure and labeled with Cy5 dye (CAT # KIT0619, Applied Biosystems).
|
| |
|
| Channel 2 |
| Source name |
Universal Mouse Reference
|
| Organism |
Mus musculus |
| Characteristics |
sample type: Universal Mouse Reference
|
| Growth protocol |
NMuMG-NT2197 cells were generated, cultured, and transfected with a pMSCV/hygro expression vector (Clontech) expressing a wildtype ShcA cDNA or a ShcA mutant harbouring an arginine to glutamine substitution at amino acid 175 (R175Q)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA (1-2.5ng) from microdissected material was subjected to two rounds of linear amplification using a RiboAmp HSPlus Amplification Kit (Cat. #: KIT0525, Applied Biosystems) following the manufacturer’s protocol.
|
| Label |
Cy5
|
| Label protocol |
The profiles of resulting amplified RNA (aRNA) were assessed using a 2100 Bioanalyzer (Agilent Technologies). The aRNA samples (5μg) were conjugated to Cy3 dye using an Arcturus Turbo Labeling Kit (Cat. #: KIT0609, Applied Biosystems). Universal human reference RNA (Stratagene) was amplified using the same procedure and labeled with Cy5 dye (CAT # KIT0619, Applied Biosystems).
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Images were quantified using Agilent Feature Extraction Software
|
| Description |
R175Q
|
| Data processing |
Microarray data were extracted using Feature Extraction Software (Agilent Technologies). The arrays were normalized using normexp background correction, loess within array and quantile between array normalization.
|
| |
|
| Submission date |
Oct 19, 2012 |
| Last update date |
Aug 07, 2013 |
| Contact name |
Sean Cory |
| E-mail(s) |
[email protected]
|
| Organization name |
McGill
|
| Department |
Goodman Cancer centre
|
| Lab |
Hallett
|
| Street address |
3649 Promenade Sir William Osler
|
| City |
Montreal |
| State/province |
QC |
| ZIP/Postal code |
H3G 0B1 |
| Country |
Canada |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE41718 |
The ShcA PTB Domain Functions as a Biological Sensor of Phospho-tyrosine Signaling During Breast Cancer Progression |
|
