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Ionic® FFPE to Pure RNA
Automated purification of RNA, including mRNA and miRNA, with minimal hands-on time
VIEW WEBINAR VIEW BROCHUREThe Ionic® FFPE to Pure RNA Kit provides automated purification of RNA, including mRNA and miRNA, from FFPE tissue samples with less hands-on time than conventional bead and column-based methods
The Ionic Purification System from Purigen Biosystems uses isotachophoresis (ITP) to extract, purify, and concentrate genomic DNA and RNA from cells and FFPE. Since nucleic acids remain in their native form, not denatured or dehydrated, the Ionic system is ideal for challenging samples with limited starting material or low-quality material.
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Get pure and native RNA, both mRNA and miRNA, from FFPE samples with higher yield and quality in half the overall number of hands-on steps per FFPE sample.
FIGURE 1: Replicate 10 µm sections from 17 FFPE tissue blocks were extracted and purified by both the Ionic FFPE to Pure RNA Kit and a market-leading column-based RNA extraction kit. The extracted and purified material from each kit was measured using a Qubit fluorometer and a Qubit High Sensitivity RNA assay. The average yield across the replicate samples processed by each method for each block is plotted as a bar graph with error bars representing the standard deviation within each population.
FIGURE 2: Libraries were prepared from RNA extracted and purified from replicate samples using the Ionic FFPE to Pure RNA Kit and a market-leading column-based RNA extraction kit following the AmpliSeq Illumina Immune Response Panel protocol. Libraries were then sequenced on an Illumina MiSeq sequencer. The mRNA expression results from each kit were normalized and compared by correlation analysis for each tissue type sampled. The correlation coefficient for each analysis indicates a strong correlation between the results of each kit for each tissue type sampled.
FIGURE 3: RNA from replicate sections of 8 FFPE sample blocks were purified using either the Ionic FFPE to Pure RNA Kit or a column-based miRNA extraction kit. The extracted and purified samples from each kit were analyzed by qPCR and the Applied Biosystems TaqMan Advanced miRNA Assays for miR-16 and miR-21. The concentration of the target miRNA represented in each sample was extrapolated and plotted against the tissue type of the source FFPE sample block. The Ionic system produced samples with a higher concentration of miRNA a large majority of the samples tested. For several samples the column-based miRNA extraction kit did not yield a detectable amount of miRNA.
FIGURE 4: Samples from “Colon 1” of FIGURE 3 above were analyzed for miRNA expression using the NanoString nCounter Human miRNA panel. The level of miRNA expression between replicate samples purified using the Ionic system has a Pearson correlation of 0.98. The level of miRNA expression between replicate samples purified using the Ionic system and the column-based miRNA kit has a Pearson correlation of 0.95. This analysis indicates a high reproducibility of miRNA expression across replicate samples purified using the Ionic system that is comparable to that of the column-based miRNA extraction kit.
Isotachophoresis separates and concentrates charged molecules in solution solely based on their electrophoretic mobility. Biological samples are gently lysed and added to the Purigen Ionic® Fluidic Chip which applies an electric field to isolate nucleic acid in its natural, native form.
