|
| Status |
Public on Mar 20, 2013 |
| Title |
slide 16 - Hyp SCH |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
control
|
| Organism |
Carassius auratus |
| Characteristics |
sex: female
|
| Treatment protocol |
sexually regressing (mid-june) female adult goldfish received 2 injections at 5 ul/g body weight:
1) 0.1% DMSO in 0.6% saline, and;
2) 0.6% saline
Animals were anaesthetized in MS222 5h post-injection, and tissues were harvested for RNA extraction, cDNA synthesis and microarray analysis.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Plus kit (Qiagen) including optional on-column DNase I treatment. RNA quantity was evaluated using the NanoDrop ND-1000. RNA integrity was evaluated using the BioAnalyzer (Agilent); RIN for each sample was > 8.4.
|
| Label |
Cy3
|
| Label protocol |
Array 900MPX™ Expression Array Detection Kit (Genisphere; see http://www.genisphere.com/array_detection_protocols.html)
|
| |
|
| Channel 2 |
| Source name |
SCH 23390 40 ug/g
|
| Organism |
Carassius auratus |
| Characteristics |
sex: female
|
| Treatment protocol |
sexually regressing (mid-june) female adult goldfish received 2 injections at 5 ul/g body weight:
1) 40 ug/g SCH 233090 0.1% DMSO in 0.6% saline, and;
2) 0.6% saline
Animals were anaesthetized in MS222 5h post-injection, and tissues were harvested for RNA extraction, cDNA synthesis and microarray analysis.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini Plus kit (Qiagen) including optional on-column DNase I treatment. RNA quantity was evaluated using the NanoDrop ND-1000. RNA integrity was evaluated using the BioAnalyzer (Agilent); RIN for each sample was > 8.4.
|
| Label |
Cy5
|
| Label protocol |
Array 900MPX™ Expression Array Detection Kit (Genisphere; see http://www.genisphere.com/array_detection_protocols.html)
|
| |
|
| |
| Hybridization protocol |
Array 900MPX™ Expression Array Detection Kit (Genisphere; see http://www.genisphere.com/array_detection_protocols.html)
|
| Scan protocol |
Microarrays were scanned at full speed 10 micron resolution using the ScanArray 5000 XL system (Packard Biosciences/PerkinElmer) using both red and blue lasers. Images were obtained with ScanArray Express software using automatic calibration sensitivity varying PMT gain (PMT starting at 65% for Cy5 and 70% for Cy3) with fixed laser power at 80% and the target intensity set for 90%.
|
| Description |
Slide #16
|
| Data processing |
Microarray images were analyzed using QuantArray (Packard Biosciences/PerkinElmer) and raw signal intensity values were obtained for duplicate spots of genes. Normalization was performed using LOWESS and GPA methods.
|
| |
|
| Submission date |
Apr 16, 2009 |
| Last update date |
Mar 20, 2013 |
| Contact name |
Jason T. Popesku |
| E-mail(s) |
jpope078@uottawa.ca
|
| Organization name |
University of Ottawa
|
| Department |
Biology
|
| Lab |
Trudeau
|
| Street address |
20 Marie Curie
|
| City |
Ottawa |
| State/province |
ON |
| ZIP/Postal code |
K1N 6N5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL7056 |
| Series (2) |
| GSE15708 |
AURATUS goldfish environmental genomics projects: SCH 23390 in the hypothalamus |
| GSE15763 |
AURATUS goldfish environmental genomics projects: dopamine antagonists |
|
