cell line: Abrams cell type: Canine OS cell line treatment: naïve
Extracted molecule
total RNA
Extraction protocol
1mL of TRI Reagent (Zymo Research, California, USA) was added to each well of cells to harvest cells for RNA extraction. the Direct-zol RNA miniprep extraction kit (product# R2051, Zymo Research, California, USA) was utilized for RNA extraction by following the manufacturer’s protocol.
Label
SYBR Green
Label protocol
Qiagen RT2 First Strand kit was utilized to synthesize 200ng cDNA in accordance with the manufacturer’s protocol (Qiagen, Hilden, Germany).
Hybridization protocol
n/a
Scan protocol
n/a
Description
Control
Data processing
Arrays were analysed with an Applied Biosystems 7500 Fast Real-Time PCR System (Thermo Fisher Scientific, Massachusetts, USA). The cycle threshold (Ct) values acquired from array analysis were analysed using the GeneGlobe RT2 Profiler PCR Data Analysis tool (Qiagen, Hilden, Germany) which calculated fold change relative to untreated cells for each evaluated OS cell line, via the ∆∆Ct methodology. The house keeping gene GAPDH was selected for normalization because treatment did not seem to alter expression. Ct values were normalized to GAPDH (matrix normalized template) and fold change was computed relative to untreated control cells (fold change template)
