Application Guide - QIAxcel Advanced

QIAxcel Advanced Application Guide 10/2016 11 0.6 0.9 0.3 1.2 1.5 1.8 0 (RFU x 1EO) 0.6 0.9 0.3 1.2 1.5 1.8 0 (RFU x 1EO) B Materials and methods Total RNA isolation from Schizosaccharomyces pombe (fission yeast) was performed according to standard methods. The protocols described below apply to RNA isolated by different methods and from different organisms. RNA samples were prepared for capillary electrophoresis according to the protocol in the QIAxcel RNA Handbook: A 1 µl aliquot of the RNA eluate was mixed with 1 µl RNA denaturing buffer, heated at 70°C for 2 minutes, and cooled in ice-cold water. Sample volume was adjusted to 10 µl with QX RNA Dilution Buffer, and samples were subsequently analyzed on the QIAxcel system using the QIAxcel RNA QC Kit v2.0 and the CM-RNA method. cRNA generation and labeling Synthesis of double-stranded cDNA, amplification to cRNA, and labeling were performed as described (1). After amplification, cRNA was normalized to 600 ng/µl and analyzed using the QIAxcel system. After quality control, the cRNA was labeled with Cy3 or Cy5 dyes and then hybridized to self-spotted or commercial microarrays. Figure 2. Streamlined RNA analysis using the QIAxcel system. Total RNA purified from Schizosaccharomyces pombe. Results presented as A a gel image and B a superimposed electropherogram view. Figure 3. Screenshot showing 28S/18S rRNA peak ratios calculated from samples shown in Figure 2A. A

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