The specificities of RT-PCR products were confirmed by both a single dissociation curve of the product and a single band with corresponding molecular weight revealed by agarose gel electrophoresis. In addition to the non-template control and negative RT control, an anatomical negative control was also performed by using the corpus callosum in the same brain, to verify the specificity of the quantitative PCR. No signal above threshold of OX was detected by quantitative PCR in any of these controls.
