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Isolate It All: siRNA miRNA ,,, Total RNA Native Protein

timized procedures; Ambion's mirVana miRNA Isolation Kit was specifically optimized for efficient isolation of RNA species smaller than 200 nt [1]. To develop the mirVana PARIS Kit, this technology was adapted and combined with the advantages of the PARIS Kit, the first commercially available kit designed to isolate both RNA and native protein from the same sample [2]. The result is a versatile kit for quantitative recovery of native protein and all RNA species, including small RNAs. A summary of each kit's specification is presented in Figure 1, and specific examples are provided below.


Figure 1. Which Kit to Choose?


Recover High Quality Total RNA
To assess the quality of total RNA isolated with the mirVana PARIS Kit, 1 x 106 HeLa cells were homogenized in the Cell Disruption Buffer provided with the kit. Because the homogenization is performed quickly on ice and in the presence of detergent, both protein and RNA can be recovered from the lysate. A fraction of the lysate can be used directly to analyze protein expression (e.g. Western blot, 2D gel electrophoresis) or for assays that require native protein (e.g. functional assay, reporter activity). For RNA isolation, the HeLa cell lysate was immediately mixed with an equal volume of 2X Denaturing Solution. This solution contains a high concentration of chaotropic denaturant
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