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Improve RNA Isolation for Array Analysis and More


In yeast, gene expression patterns can be affected during pretreatments used to disrupt the yeast cells prior to RNA isolation. The RiboPure-Yeast RNA Isolation Kit uses no enzymatic pretreatments and thus does not alter expression profiles. RiboPure-Yeast combines a simple and efficient glass bead/one step phenol mediated disruption using Ambion's Vortex Adapter followed by a glass filter based RNA purification step. After DNase treatment with the included DNA-free Reagents, the resulting RNA is exceptionally pure (figure 1) containing little to no genomic DNA (<0.05%). The kit has been used at Ambion to isolate high quality RNA from Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, and Candida albicans. The kit provides the highest yield of any method tested from each of these species (Figure 2). The RiboPure-Yeast RNA Isolation Kit contains sufficient reagents for 50 RNA isolations each from 107 up to 3 x 108 yeast cells.

Figure 1. Yeast Total RNA Isolated Using the RiboPure-Yeast Kit. P. pastoris, S. cerevisiae RNA, C. albicans, and S. pombe RNA, isolated using the RiboPure-Yeast Kit and treated with DNase, was run on an Agilent bioanalyzer chip. The ratio of 28S:18S rRNA is given for each sample.

Figure 2. RNA Yields From 3 Yeast Species Using Ambion's RiboPure-Yeast or Competitors' Kits. RNA was isolated from the specified number of yeast cells using the manufacturer's protocol. Following purification,
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