|
Status |
Public on Mar 20, 2013 |
Title |
slide 47 - Tel SKF |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
control
|
Organism |
Carassius auratus |
Characteristics |
Sexually mature (mid-May) 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 Telencephalon tissue was extracted 5h post-injection.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated with the TRIzol reagent (Invitrogen, Burlington, ON, Canada) per the manufacturer's protocol. Samples were treated with DNase on-column in an RNeasy Mini Plus kit (Qiagen, Mississauga, ON, Canada). RNA quantity was evaluated using the NanoDrop. 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 |
SKF 38393 40 ug/g
|
Organism |
Carassius auratus |
Characteristics |
Sexually mature (mid-May) female adult goldfish received 2 injections at 5 uL/g body weight: 1) SKF 38393 40 ug/g in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Telencephalon tissue was extracted 5h post-injection.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated with the TRIzol reagent (Invitrogen, Burlington, ON, Canada) per the manufacturer's protocol. Samples were treated with DNase on-column in an RNeasy Mini Plus kit (Qiagen, Mississauga, ON, Canada). RNA quantity was evaluated using the NanoDrop. 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 |
Sexually mature female goldfish received 2 injections at 5 uL/g body weight: 1) 0.1% DMSO and 0.6% saline (control) or SKF 38393 40 ug/g in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Animals were anaesthetized in MS222, and telencephalon tissues were harvested for RNA extraction, cDNA synthesis and microarray analysis. Slide #47; dye-swap of slide #30
|
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 |
Jan 27, 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 |
GPL3735 |
Series (2) |
GSE14608 |
AURATUS goldfish environmental genomics projects: SKF 38393 in the telencephalon |
GSE15855 |
AURATUS goldfish environmental genomics projects: dopamine agonists |
|