Expression profiles of MicroRNA and SiRNA of Arabidopsis thaliana Col-0 and transgenic plants

Expression profiles of MicroRNA and SiRNA of Arabidopsis thaliana Col-0 and transgenic plants with constitutive expression of the chimeric receptors NRG1 grown at different temperature To reveal the underlying molecular mechanism of de-cosuppression with memory by high temperature in Arabidopsis, we performed the expression profiles of microRNA and SiRNA in transgenic plants with constitutive expression of the chimeric receptors NRG1 and wide type Col-0 grown at different temperature using the Custom LC Sciences Arabidopsis microRNA and SiRNA array. Keywords: high temperature, de-cosuppression, MicroRNA, SiRNA Total RNA was prepared from wild type and NRG1 plants grown at 22ºC and 30ºC for 4 weeks with TRIzol reagent according to the manufacturer’s protocol (Invitrogen). Small RNAs (<300nt) were isolated using a YM-100 Microcon centrifugal filter (Millipore), and 3’-extended with a poly(A) tail for later fluorescent dye staining. Two different tags were used for the two RNA samples labeled with Cy3 or Cy5 fluorescent dyes in dual-sample experiments. Also dye-swap experiments were carried out to control for labeling bias. Hybridization was performed overnight on a µParafloTM microfluidic chip. Each miRNA and siRNA had 6 and 3 repeat prints, respectively. Six replicate arrays were performed with six independent pairs of RNA samples. After hybridization, array images were obtained using an Axon GenePix 4000B laser scanner and digitized using the Array-Pro image software (Media Cybernetics). The data were corrected by subtracting the background and normalized using the LOWESS (locally weighted regression) method. All data calculations were performed by LC Science (Houston, TX).