|
| Status |
Public on Oct 18, 2010 |
| Title |
Norm.1.col.Ref1.2 (Exiqon) |
| Sample type |
RNA |
| |
|
| Source name |
Total RNA with small fraction
|
| Organism |
Mus musculus |
| Characteristics |
sample: Reference 1
tissue: pool of equal amounts of total RNA from mouse testicle, ovary and 10-12 day embryo.
sample type: Reference RNA was developed from FirstChoice® mouse Total RNA including the small fraction (Catalogue # AM7800-AM7828, Ambion, Streetsville, ON)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA containing the small fraction was isolated from each sample using the miRVana miRNA Isolation Kit (Ambion), Streetsville, ON, Canada). Quality was confirmed using the Agilent 2100 Bioanalyzer and RNA Nano 6000 chips.
|
| Label |
Hy3
|
| Label protocol |
One microgram of each mouse Reference RNA sample was labelled with a Hy5 fluorophore (Exiqon, Woburn, MA, USA) and co-hybridised with 1 g of the other Reference RNA standard in Hy3. Labelling reactions were performed using Exiqon’s miRCURY LNA miRNA power labelling kit, according to the manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Hybridisations were performed in SureHyb chambers (Agilent) for 16 hours at 56ºC. Slides were washed according to manufacturer’s instructions
|
| Scan protocol |
Arrays were scanned at a resolution of 5 mm using an Agilent G2505B scanner. Data were acquired using Agilent Feature Extraction software version 9.5.3.1, using an FE protocol available on-demand from Exiqon.
|
| Description |
Hy3: CHIP # 2 flipUS22502706_13974287_S01_Exiqon1.txt
|
| Data processing |
Data were extracted from scanned images using Agilent Feature Extraction software version 9.5.3.1. Median signal intensities were lowess normalized for the 2 colour analysis and cyclic lowess normalized for the one colour analysis. Technical replicates for each miRNA level were averaged using the median. Data was flagged if the raw signal intensity was less than the mean plus three standard deviations of the negative controls or if the median of the technical replicates was flagged to be in the background.
|
| |
|
| Submission date |
Dec 29, 2009 |
| Last update date |
Oct 18, 2010 |
| Contact name |
Andrew Williams |
| Organization name |
Health Canada
|
| Street address |
50 Columbine
|
| City |
Ottawa |
| State/province |
ON |
| ZIP/Postal code |
K1A OK9 |
| Country |
Canada |
| |
|
| Platform ID |
GPL7723 |
| Series (1) |
| GSE19669 |
Cross-platform correlation analysis of global microRNA expression technologies |
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