|
| Status |
Public on Nov 12, 2010 |
| Title |
Control #1: mock transfection of HCT116 dicer- cell line, 24 h timepoint |
| Sample type |
RNA |
| |
|
| Source name |
HCT-116 dicer- cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HCT-116 dicer- cells
conditions: mock transfection
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNAeasy
|
| Label |
Biotin
|
| 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 deg 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.
|
| 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).
|
| |
|
| Submission date |
Nov 08, 2010 |
| Last update date |
Nov 12, 2010 |
| Contact name |
Lee Lim |
| E-mail(s) |
lee_lim@merck.com
|
| Phone |
206-802-7323
|
| Organization name |
Rosetta Inpharmatics
|
| Street address |
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98109 |
| Country |
USA |
| |
|
| Platform ID |
GPL10379 |
| Series (1) |
| GSE25224 |
Persistence of seed-based activity following microRNA segmentation |
|
