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Unmask Blood RNA for Gene Expression Profiling

> Electropherogram of Amplified RNA from Human Whole Blood. Total RNA was isolated from whole blood using the RiboPure-Blood Kit (Ambion). Total RNA (5 g) was treated with the GLOBINclear Kit (Ambion). MessageAmp II aRNA Amplification Kit (Ambion) was used to linearly amplify 2 g total RNA or RNA enriched by the GLOBINclear Kit. The resulting aRNA was resolved on an Agilent 2100 bioanalyzer RNA LabChip assay to monitor quality and quantity. Note the complete removal of the abundant "globin spike" in the GLOBINclear-treated RNA that is present in the whole blood aRNA sample.


Successful Gene Expression Profiling

To further test the specificity of globin mRNA reduction, synthetic globin transcripts were added to HeLa cell RNA, and samples were processed or left unprocessed with the GLOBINclear Kit (Figure 2). In microarray analyses, the GLOBINclear protocol did not alter the profiles from HeLa RNA preparations. However, the GLOBINclear process was able to rescue the decreased gene detection observed with RNA samples containing high levels of exogenous globin transcripts.

Figure 2. Percent Present Calls for Samples Treated or Untreated with GLOBINclear-Human Globin mRNA Removal Kit. In a control experiment, the GLOBINclear Kit (Ambion) did not significantly alter the Percent Present Call results from RNA samples derived from a cultured cell line (HeLa cells) that does not express high levels of globin (left two bars). In contrast, the GLOBINclear Kit rescued the decreased gene detection observed when exogenous globin transcripts were spiked into the HeLa RNA samples. Total RNA was isolated from human blood
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