Stool Total RNA Purification Kit Dx A convenient and rapid method to purify total RNA from small amounts of stool samples for in vitro diagnostic use Note: This product is not available for sale in the United StatesResearch version available here Powered by Bioz Stool Total RNA Purification Kit Dx A convenient and rapid method to purify total RNA from small amounts of stool samples for in vitro diagnostic use Price USD$499.00 SKU Dx49500 Format Spin Column Size 50 Preps Quantity Overview Details Documentation Citations Overview CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746. Ideal for use in in vitro diagnostic workflows Simultaneous isolation of both host RNA and microbial RNA (universal protocol) Isolate full diversity of RNA from large RNA down to small and microRNAs Eliminates PCR inhibitors including humic acids High quality RNA for sensitive downstream applications This kit provides a convenient and rapid method to purify total RNA from small amounts of stool samples. All types of stool samples can be processed with this kit, including animal fecal samples, manure and samples collected using Norgen's Stool Nucleic Acid Collection and Transport Devices Dx (Cat. Dx45660). The kit removes all traces of humic acids using rapid and simple spin column procedures. Bead tubes are also provided for effective homogenization of stool. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. Both host and microbial RNA is recovered. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR and reverse transcription PCR for gene expression analysis. The procedure can be completed in approximately 30 minutes. NOTE: This product is not available for sale in the United States. Details Supporting Data Previous Figure 1. High yields of stool total RNA were purified from 200 mg of human stool using Norgen's Stool RNA Purification Kit Dx and a leading competitor's kit, in triplicate. For analysis, 7.5 µL of each 75 µL elution was loaded on a 1.2 % 1x MOPS formaldehyde-agarose gel. Norgen's kit was found to have a higher yield of RNA, isolated from 200 mg of human infant stool. Figure 2. Yield and Quality of Purified Stool RNA. Total RNA was isolated from 200 mg of human stool using Norgen's Stool RNA Purification Kit Dx and a leading competitor kit. Comparisons were then made based on yield, and A260:A280/A260:A230 ratios measured using the NanoVue Plus™. In Panel A it can be seen that both kits isolated RNA with high A260:A280 ratios (all samples were found to be above 1.8 and below 2.2). In Panel B, Norgen's kit was found to isolate RNA with a high A260:A230 (with all samples once again falling in the 1.8-2.2 range). The competitor kit, however, was found to isolate RNA with extremely low A260:A230 ratios, with none of the samples displaying a A260:A230 ratio higher than 0.20. The results in Panel C are in agreement with the gel photo from Figure 1, and Norgen's kit was found to isolate higher amounts of RNA, with an average yield of 14.58 µg, compared to the competitor’s average of 12.26 µg. Figure 3. Detection of Bacterial Stool RNA using 16S Primers. Total stool RNA was isolated from 200 mg human stool samples using Norgen's Stool RNA Purification Kit Dx and a leading competitor’s kit. Five microliters of purified RNA was used in a 20 µL reverse-transcription reaction using Invitrogen's Superscript III system with 16S reverse primers. The cDNA generated was then used in a qPCR reaction involving Norgen's 2X PCR Mastermix spiked with SYBR green (Bio-Rad), using 0.3µM of primers against bacterial 16S. As can be seen in the amplification plot, Norgen's kit outperformed the leading competitor's kit by on average 1.5 Ct values. This indicates that Norgen isolated higher quality and yields of RNA from stool, that can be used in an array of downstream applications. Next Click for expanded view Kit Specifications Maximum Stool Input 200 mg Type of Stool Processed Frozen and fresh stool Maximum Column Binding Capacity 50 μg Maximum Column Loading Volume 650 μL Time to Complete 10 Purifications 30 minutes Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. Component Cat. Dx49500 (50 preps) Lysis Solution 60 mL Wash Solution 22 mL Elution Buffer 6 mL Bead Tubes 50 Spin Columns 50 Collection Tubes 50 Elution Tubes (1.7 mL) 50 Product Insert 1 Documentation PROTOCOLS (Dx49500) Stool Total RNA Purification Kit Dx - Protocol (50 prep) APPLICATION NOTES Stool Total RNA Purification Kit Purification and Detection of Bacterial RNA From Preserved Stool Samples - Application Note FLYERS Flyer Citations Title The role of whole-grain barley on human fecal microbiota and metabolome Journal Applied and Environmental Microbiology. 2015. Authors Maria De Angelis, Eustacchio Montemurno, Lucia Vannini, Carmela Cosola, Noemi Cavallo, Giorgia Gozzi, Valentina Maranzano, Raffaella Di Cagno, Marco Gobbettia and Loreto Gesualdo Title Microbiota and metabolome of un-started and started Greek-type fermentation of Bella di Cerignola table olives Journal Food Microbiology. 2015. Authors Maria De Angelis, Daniela Campanella, Lucrezia Cosmai, Carmine Summo, Carlo Giuseppe Rizzello, Francesco Caponio Title Stool metatranscriptomics: A technical guideline for mRNA stabilisation and isolation Journal BMC Genomics. 2015. Authors Michael Reck, Jürgen Tomasch, Zhiluo Deng, Michael Jarek, Peter Husemann, Irene Wagner-Döbler Title Fecal Microbiota in Healthy Subjects Following Omnivore, Vegetarian and Vegan Diets: Culturable Populations and rRNA DGGE Profiling Journal PLOS ONE. 2015. Authors Ilario Ferrocino, Raffaella Di Cagno, Maria De Angelis, Silvia Turroni, Lucia Vannini, Elena Bancalari, Kalliopi Rantsiou, Gianluigi Cardinali, Erasmo Neviani, Luca Cocolin Title Modulation of the Intestinal Microbiota Is Associated with Lower Plasma Cholesterol and Weight Gain in Hamsters Fed Chardonnay Grape Seed Flour. Journal Journal of Agriculture and Food Chemistry. 2015. Authors Hyunsook Kim, Dong-Hyeon Kim, Kun-ho Seo, Jung-Whan Chon, Seung-Yeol Nah, Glenn E. Bartley, Torey Arvik, Rebecca Lipson, and Wallace Yokoyama. Title Microbiota and Metabolome Associated with Immunoglobulin A Nephropathy (IgAN). Journal Microbiota and Metabolome of IgA Nephropathy Patients. 2014. Authors Maria De Angelis, Eustacchio Montemurno, Maria Piccolo, Lucia Vannini, Gabriella Lauriero, Valentina Maranzano, Giorgia Gozzi, Diana Serrazanetti, Giuseppe Dalfino, Marco Gobbetti, Loreto Gesualdo. 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Stool Total RNA Purification Kit Dx A convenient and rapid method to purify total RNA from small amounts of stool samples for in vitro diagnostic use Note: This product is not available for sale in the United StatesResearch version available here Powered by Bioz Stool Total RNA Purification Kit Dx A convenient and rapid method to purify total RNA from small amounts of stool samples for in vitro diagnostic use Price USD$499.00 SKU Dx49500 Format Spin Column Size 50 Preps Quantity