|
| Status |
Public on May 03, 2012 |
| Title |
JH033 xenograft GSI_JH033B Sensitive to MRK-003 |
| Sample type |
RNA |
| |
|
| Source name |
pancreatic ductal adenocarcinoma cell lines
|
| Organism |
Homo sapiens |
| Characteristics |
sample: GSI_JH033B
donor cell line: JH033
host: immunocompromised Athymic Nude-Foxn1nu mouse
phenotype: Sensitive to MRK-003
tissue: Pancreatic ductal adenocarcinoma tumor xenograft
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples are processed in parallel in 96-well plates to minimize potential variation. Reaction purification is achieved using magnetic binding beads for cDNA and Qiagen RNeasy kits for cRNA purification.
|
| Label |
Biotin ining process that provides the measurable signal.
|
| Label protocol |
The final in vitro transcription incorporates biotin moieties that are later labeled with phycoerythrin. After amplification, samples are put through a controlled fragmentation to improve hybridization sensitivity and consistency. The labeled molecules are biotinylated-cRNA.
|
| |
|
| Hybridization protocol |
GeneChip microarrays are loaded with the fragmented target sample/hybridization buffer mix using standard manual techniques. Arrays are hybridized for 18 hours at 45?C with vigorous mixing. Unbound sample is removed and staining is accomplished through the binding of streptavidin conjugated phycoerythrin to the hybridized target. Excess label is removed. Washing and staining steps are performed by the Affymetrix FS450 fluidics station using standard protocols.
|
| Scan protocol |
Arrays are scanned using a GeneChip Scanner 3000 7G with a 48 array autoloader. The scanner maintains the optimal temperature for the arrays prior to and during scanning.
|
| Description |
We used pancreatic ductal adenocarcinoma cell lines to determine whether pharmacological targeting of the Notch pathway could inhibit pancreatic tumor growth and potentiate gemcitabine sensitivity.
|
| Data processing |
Data were loaded into the Rosetta Resolver? system (Rosetta Biosoftware, Seattle, WA) and processed using the RMA algorithm (http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=PubMed&list_uids=12582260). Intensities were calculated based on RMA, a Rosetta-developed error, and a MAS-5 p-value. The Rosetta-developed error is used in the calculation of xdev for the ratio p-values as described in section 2.2 (http://bioinformatics.oxfordjournals.org/cgi/content/full/22/9/1111).
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| |
|
| Submission date |
Apr 29, 2012 |
| Last update date |
May 03, 2012 |
| Contact name |
Oscar Puig |
| E-mail(s) |
oscar_puig@merck.com
|
| Organization name |
Merck Research Laboratories
|
| Department |
Molecular Profiling Research Informatics
|
| Street address |
126 East Lincoln Ave
|
| City |
Rahway |
| State/province |
NJ |
| ZIP/Postal code |
07065 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE37645 |
The gamma secretase inhibitor MRK-003 attenuates pancreatic cancer growth in preclinical models |
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