|
Status |
Public on May 02, 2013 |
Title |
Control Day 3, Sample.40 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
Heart tissue, control, day 3
|
Organism |
Mus musculus |
Characteristics |
strain: C57BL/6 gender: female tissue: heart treatment: vehicle control dose: 0 ug time point: day 3 sampleID: 358
|
Treatment protocol |
Female C57BL/6 mice (Taconic, Denmark) were acquired at 5-6 weeks of age and given food (Altromin no 1324; Christian Petersen, Denmark) and water ad libitum. Printex 90 was suspended by sonication in 0.9% NaCl MilliQ water containing 10% v/v acellular bronchial alveolar lavage fluid from C57BL/6 mice. A 40 µl suspension was instilled followed by 150 μl air.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using the mirVana Isolation kit (Ambion, Canada).
|
Label |
Cy5
|
Label protocol |
Individual 200 ng aliquots of RNA from each sample were amplified and labelled using the Low RNA Input Fluorescent Linear Amplification Kit (Agilent, (5184-3523)).
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|
|
Channel 2 |
Source name |
Universal Mouse Reference RNA
|
Organism |
Mus musculus |
Characteristics |
sample type: reference
|
Extracted molecule |
total RNA |
Extraction protocol |
Material was purchased as total RNA (Universal Mouse Reference RNA) from Stratagene (CA, USA).
|
Label |
Cy3
|
Label protocol |
Individual 200 ng aliquots of RNA from each sample were amplified and labelled using the Low RNA Input Fluorescent Linear Amplification Kit (Agilent, (5184-3523)).
|
|
|
|
Hybridization protocol |
Agilent Mouse G4852A Microarrays (containing approximately 44,000 probes) were hybridized with 300 ng Cy5-labelled heart RNA and 300 ng Cy3-labelled Universal Mouse Reference RNA (Stratagene, CA, USA), used as a common reference on all arrays. Arrays were incubated for 17 hours at 65 ºC in Agilent hybridization solution and washed according to manufacturer’s instructions.
|
Scan protocol |
Arrays were scanned on an Agilent G2505B scanner. Data were acquired using Agilent Feature Extraction software version 10.7.3.1.
|
Description |
Sample.40
|
Data processing |
The log2 of the ratio (sample(Cy5)/reference(Cy3)), using the median signal intensities, were normalized using lowess using the transform.madata function in the maanova library in R.
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|
|
Submission date |
May 09, 2012 |
Last update date |
May 02, 2013 |
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 |
GPL13912 |
Series (2) |
GSE37887 |
Cardiac gene expression (random section containing both ventricles) in C57BL/6 mice intratracheally instilled with Printex 90 carbon black nanoparticles (set 2) |
GSE37891 |
Cardiac gene and miRNA expression (random section containing both ventricles) in C57BL/6 mice intratracheally instilled with Printex 90 carbon black nanoparticles |
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