Name: Stool Nucleic Acid Isolation Kit
Brand: Norgen Biotek Corp.
SKU: 45600
Price: 759.000000 USD
Availability: InStock

Features and Benefits

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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Details

Supporting Data

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.

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.

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

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

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	Comparison of intestinal bacterial communities in asymptomatic and diseased Asian seabass (Lates calcarifer) with chronic enteritis and mixed bacterial infections
Citation	Aquaculture 2023.
Authors	Xian Zhe Chew, Susan Gibson-Kueh, Dean R. Jerry and Xueyan Shen

Title	Gut microbiome dysbiosis is associated with host genetics in the Norwegian Lundehund
Citation	Frontiers in Microbiology 2023.
Authors	Claudia Melis, Anna Maria Billing, Per-Arvid Wold, William Basil Ludington

Title	Gut Microbiota Composition in the First and Third Trimester of Pregnancy among Malay Women is Associated with Body Mass Index: A Pilot Study
Citation	The Malaysian Journal of Medical Sciences : MJMS 2023.
Authors	Bahiyah Abdullah, Mohd Yusri Idorus, Suzanna Daud, Shafiq Aazmi, Thanikasalam Kathiresan Pillai, and Zaini Mohd Zain

Title	Positive modulation of a new reconstructed human gut microbiota by Maitake extract helpfully boosts the intestinal environment in vitro
Citation	Plos One 2023.
Authors	Alessandra De Giani,Federica Perillo,Alberto Baeri,Margherita Finazzi,Federica Facciotti,Patrizia Di Gennaro

Title	Sex-specific effects of voluntary wheel running on behavior and the gut microbiota-immune-brain axis in mice
Citation	Brain Behav Immun Health. 2023.
Authors	Zoë A.P. Williams, Joanna Kasia Szyszkowicz, Natasha Osborne, Bshaier Allehyany, Christophe Nadon, Maryann Chinonye Udechukwu, Ana Santos and Marie-Claude Audet

Title	Sympathovagal quotient and resting-state functional connectivity of control networks are related to gut Ruminococcaceae abundance in healthy men
Citation	Psychoneuroendocrinology 2023.
Authors	Ana L. Miranda-Angulo a, Juan D. Sánchez-López a, Daniel A. Vargas-Tejada a, Hawkins-Caicedo Valentina a, Juan C. Calderón a, Jaime Gallo-Villegas b c, Juan F. Alzate-Restrepo d, Jazmin X. Suarez-Revelo e, Gabriel Castrillón e f

Stool Nucleic Acid Isolation Kit