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.
Stool Nucleic Acid Isolation Kit
A convenient and rapid method to isolate total DNA and RNA from fresh, frozen and preserved stool samples.
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.
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Supporting Data
Kit Specifications
|
|
Maximum Stool Input
|
200 mg
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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.
Component | Cat. 45600 (50 preps) |
---|---|
Lysis Buffer C | 60 mL |
Binding Buffer E | 6 mL |
Wash Solution A | 38 mL |
Elution Buffer E | 6 mL |
Bead Tube | 50 |
Spin Columns | 50 |
Collection Tubes | 50 |
Elution Tubes (1.7 mL) | 50 |
Product Insert | 1 |
Documentation
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
FAQs
Spin Column
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.
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 |