Stool Nucleic Acid Isolation Kit A convenient and rapid method to isolate total DNA and RNA from fresh, frozen and preserved stool samples. For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Stool Nucleic Acid Isolation Kit A convenient and rapid method to isolate total DNA and RNA from fresh, frozen and preserved stool samples. Price USD$759.00 SKU 45600 Format Spin Column Size 50 Preps Quantity Overview Details Documentation FAQs Citations Overview Simultaneous isolation of both host and microbial DNA and RNA Eliminates PCR inhibitors including humic acids High quality total RNA and DNA for sensitive downstream applications Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix This kit provides a rapid spin column method to isolate total DNA and RNA from fresh or frozen stool samples, as well as from stool collected using Norgen's Stool Nucleic Acid Collection and Preservation Tubes. The universal protocol conveniently allows for the isolation of total genomic DNA and total RNA from all the various microorganisms and host cells found in the stool sample simultaneously. The kit removes all traces of humic acids and provides consistent, high yields of RNA and DNA. The purified DNA and RNA are of the highest quality and are fully compatible with downstream PCR applications, as all humic acid substances and other PCR inhibitors are removed during the isolation. Details Supporting Data Previous Figure 1. Total RNA Profile from Different Stool Samples. Total RNA and DNA were isolated from 6 different 200 mg human stool samples using Norgen's Stool Nucleic Acid Isolation Kit. For analysis, 7.5 µL from each 75 µL elution were loaded on 1.2 % MOPS agrarose gel. All six sample showed a good RNA integrity and total RNA profile that includes large and small RNA. Figure 2. Detection of Bacterial RNA from Stool Samples. Real-time one step RT-PCR analysis (SYBR Green) was carried out to detect the bacterial 16s rRNA expression from six stool samples isolated using Norgen's Stool Nucleic Acid Isolation Kit. The RNA template was treated with DNase I and concentrated with Norgen's RNA Clean-Up and Concentration Kit (Cat# 23600, 43200). No PCR inhibition was observed from 300 ng of total RNA in 20 µL PCR reaction. Black diamond: negative control, Blue diamond: six stool RNA samples. Figure 3. Isolation of Genomic DNA from Human Stool Samples. Total RNA and DNA was isolated from 6 different 200 mg human stool samples using Norgen's Stool Nucleic Acid Isolation Kit. For analysis, 10 µL of each 75 µL elution was run on a 1.2% 1xTAE agarose gel. High quality DNA was isolated from all six stool samples. M: Norgen's HighRanger 1kb DNA ladder (Cat. 11900). Figure 4. Detection of Human DNA from Stool Samples. Total RNA and DNA were isolated from two different stool samples using Norgen's Stool Nucleic Acid Isolation Kit. Two microliters of each elution was used in 20 µL of Norgen's 2X PCR Mastermix spiked with SYBR green (Bio-Rad), using 0.3 µM of primers against GAPDH. As can be seen in the amplification plot, all four samples were successfully amplified, indicating that PCR inhibitors were removed and the kit isolated high quality of DNA from stool. PCR quantification data (A) and melting curve analysis date (B) are shown. Next Click for expanded view Kit Specifications Maximum Stool Input 200 mg Type of Stool Processed Preserved, 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. ComponentCat. 45600 (50 preps)Lysis Buffer C60 mLBinding Buffer E6 mLWash Solution A38 mLElution Buffer E6 mLBead Tube50Spin Columns50Collection Tubes50Elution Tubes (1.7 mL)50Product Insert1 Documentation PROTOCOLS (45600) Stool Nucleic Acid Isolation Kit - Protocol (50 prep) APPLICATION NOTES The Isolation of High Quality Stool DNA and its Application to Quantitative Adenovirus Detection using TaqMan® Real-Time PCR Comparison of Stool DNA Isolation Methods for Bacterial and Mammalian DNA Detection Comparative Microbiome Profiles from Fecal Preservation Methods Comparison of DNA Isolated from Stool Using Norgen’s Isolation Kits Versus Competitor’s DNA Stool Mini Kit Immediate Viral Inactivation Using Norgen’s Stool Nucleic Acid Preservative POSTERS Stool Nucleic Acid Isolation Kit Effect of Stool Transportation Conditions On Microbiota Diversity - Poster RELATED BLOGS 3 Reasons why Norgen’s Industry-Leading Patented RNA Purification Technology is Best-in-Class FAQs Spin Column Why do I have poor DNA/RNA recovery? Poor DNA or RNA recovery may be caused by one or more of the following: Homogenization was incomplete. Depending on the type of stool, further vortexing with the flat bed vortex or bead beater equipment may be required. However, it is not recommended to increase the vortex time to longer than 5 minutes at maximum speed. Also, ensure that the maximum input of 200 mg of stool is not exceeded, as this may also cause incomplete homogenization. An alternative elution buffer was used. It is recommended that the Elution Buffer E supplied with this kit be used for maximum DNA and RNA recovery. Ethanol was not added to the lysate. Ensure that an equal amount of 70% ethanol is added to the lysate before binding to the column. Ethanol was not added to the Wash Solution A Ensure that 90 mL of 96-100% ethanol is added to the supplied Wash Solution A prior to use. Why is the elution not performing well in downstream applications? If the nucleic acids does not perform well in downstream applications, It may be due to one or more of the following: Eluted sample is brown. Ensure Binding Buffer E is added to the lysate and that it is incubated on ice for 10 minutes prior to spinning down the lysate. Avoid any contact with the pellet or surface residue when collecting the supernatant after the 5 minute spin during sample preparation. The nucleic acids were not washed three times with the provided Wash Solution A. Traces of salt from the binding step may remain in the sample if the column is not washed three times with the provided Wash Solution A. Salt may interfere with downstream applications, and thus must be washed from the column. Ethanol carryover. Ensure that the dry spin under the "Column Wash" procedure is performed, in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications. Binding Buffer E was not added to the lysate. Ensure that the Binding Buffer E is added to the lysate and that it is incubated on ice for 10 minutes prior to spinning down the lysate. PCR reaction conditions need to be optimized. Take steps to optimize the PCR conditions being used, including varying the amount of template, changing the source of Taq polymerase, looking into the primer design and adjusting the annealing conditions. Citations Title Positive modulation of a new reconstructed human gut microbiota by Maitake extract helpfully boosts the intestinal environment in vitro Journal Plos One. 2024. Authors Alessandra De Giani, Federica Perillo, Alberto Baeri, Margherita Finazzi, Federica Facciotti, Patrizia Di Gennaro Title Effects of Inulin-Based Prebiotics Alone or in Combination with Probiotics on Human Gut Microbiota and Markers of Immune System: A Randomized, Double-Blind, Placebo-Controlled Study in Healthy Subjects Journal Microorganisms. 2022. Authors Alessandra De Giani, Anna Sandionigi, Jessica Zampolli, Angela Michelotti, Francesco Tursi, Massimo Labra and Patrizia Di Gennaro Title Effectiveness of Multistrain Probiotic Formulation on Common Infectious Disease Symptoms and Gut Microbiota Modulation in Flu-Vaccinated Healthy Elderly Subjects Journal BioMed Research International. 2022. Authors Anna Sandionigi, Alessandra De Giani, Francesco Tursi, Angela Michelotti, Enza Cestone, Silvana Giardina, Jessica Zampolli, and Patrizia Di Gennaro Title Escherichia Coli as a Marker for Hypertriglyceridemia Journal United States Patent Application 20160168627. 2016. Authors Chou CJ, Darimont-nicolau C, Membrez M Related Products Fecal DNA Collection & Preservation Mini Tubes Stool DNA Isolation Kit Stool Total RNA Purification Kit Soil DNA Isolation Kit
Stool Nucleic Acid Isolation Kit A convenient and rapid method to isolate total DNA and RNA from fresh, frozen and preserved stool samples. For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Stool Nucleic Acid Isolation Kit A convenient and rapid method to isolate total DNA and RNA from fresh, frozen and preserved stool samples. Price USD$759.00 SKU 45600 Format Spin Column Size 50 Preps Quantity
