Isolation of total RNA from stool samples—NucleoSpin RNA Stool

The NucleoSpin RNA Stool kit is designed for efficient isolation of RNA, including small RNA species, from fresh and frozen stool samples. The kit combines NucleoZol with Lysis Buffer RST1 for chemical disruption of stool and associated microbes. The mechanical lysis is performed using NucleoSpin Bead Tubes Type A (containing ceramic beads) in combination with a mechanical disruption device. No enzymatic reactions, such as protease digestion, are required to homogenize the sample material.

Undissolved sample material and the ceramic beads are subsequently removed by a short centrifugation. A subsequent lysate clearing step consists of addition of Binding Buffer RST2 followed by a short centrifugation. The supernatant is finally cleared by passing it through a NucleoSpin Inhibitor Removal Column, which removes substances in stool samples that interfere with qPCR. Binding conditions are adjusted by adding additional Binding Buffer RST2 to the flowthrough of the NucleoSpin Inhibitor Removal Column. Next, the sample is loaded onto a NucleoSpin RNA Stool Column. Residual contaminants such as complex polysaccharides, bile salts, and other PCR inhibitors are removed by an efficient washing procedure using Buffer RST2, as well as Wash Buffers RST3, RST4, and RST5. In between the different washing steps, residual DNA is removed via an on-column DNA digestion step using rDNase. After a drying step, ready-to-use RNA can be eluted with RNase-free water.

More Less

Cat. #	Product	Size	Price
740130.10	NucleoSpin® RNA Stool	10 Preps	$115.00
The NucleoSpin RNA Stool kit is designed for efficient isolation of RNA, including small RNA species, from fresh and frozen stool samples. The kit employs silica-membrane technology in a mini spin column format, and typically yields 10–30 µg of RNA from 200-mg inputs of fresh or frozen stool. The kit workflow allows for processing of 10 samples in parallel in approximately 70 min. Cat. # 740130.10 includes sufficient quantities of reagents and materials for processing of 10 samples. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Image Data Back NucleoSpin RNA Stool procedure NucleoSpin RNA Stool procedure. Back Highest yield with NucleoSpin RNA Stool Highest yield with NucleoSpin RNA Stool. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For each sample and protocol, extractions were performed in triplicate. For each sample, NucleoSpin RNA Stool provided 20–60% higher yields compared to the competitor kits. Back Highest RNA purity and integrity with NucleoSpin RNA Stool Highest RNA purity and integrity with NucleoSpin RNA Stool. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For each sample and protocol, four extractions were performed. For each sample, NucleoSpin RNA Stool yielded RNA of the highest purity (A₂₆₀/A₂₈₀, top) and integrity (RIN, bottom) compared to the competitor kits. Back NucleoSpin RNA Stool shows best performance in qPCR NucleoSpin RNA Stool shows best performance in qPCR. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For samples processed with NucleoSpin RNA Stool and protocol "Q", RNA was eluted in 100-µl volumes, whereas eluates from protocol "Z" were each adjusted to a final volume of 100 µl to obtain comparable RNA concentrations for each protocol. For each sample and protocol, extractions were performed in triplicate. For each sample, RNA obtained with NucleoSpin RNA Stool provided better performance in qPCR compared to RNA prepared with the competitor kits.
740130.50	NucleoSpin® RNA Stool	50 Preps	$440.00
The NucleoSpin RNA Stool kit is designed for efficient isolation of RNA, including small RNA species, from fresh and frozen stool samples. The kit employs silica-membrane technology in a mini spin column format, and typically yields 10–30 µg of RNA from 200-mg inputs of fresh or frozen stool. The kit workflow allows for processing of 10 samples in parallel in approximately 70 min. Cat. # 740130.50 includes sufficient quantities of reagents and materials for processing of 50 samples. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Image Data Back NucleoSpin RNA Stool procedure NucleoSpin RNA Stool procedure. Back Highest yield with NucleoSpin RNA Stool Highest yield with NucleoSpin RNA Stool. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For each sample and protocol, extractions were performed in triplicate. For each sample, NucleoSpin RNA Stool provided 20–60% higher yields compared to the competitor kits. Back Highest RNA purity and integrity with NucleoSpin RNA Stool Highest RNA purity and integrity with NucleoSpin RNA Stool. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For each sample and protocol, four extractions were performed. For each sample, NucleoSpin RNA Stool yielded RNA of the highest purity (A₂₆₀/A₂₈₀, top) and integrity (RIN, bottom) compared to the competitor kits. Back NucleoSpin RNA Stool shows best performance in qPCR NucleoSpin RNA Stool shows best performance in qPCR. Two human stool samples were processed within 4 hours after collection using standard protocols including an on-column DNase digestion step. As indicated in the corresponding protocols, 200-mg stool samples were processed using NucleoSpin RNA Stool ("MN"), whereas 250-mg samples were processed with the competitor kits. For protocol "Q", the optional phenol-based lysis step was included. For samples processed with NucleoSpin RNA Stool and protocol "Q", RNA was eluted in 100-µl volumes, whereas eluates from protocol "Z" were each adjusted to a final volume of 100 µl to obtain comparable RNA concentrations for each protocol. For each sample and protocol, extractions were performed in triplicate. For each sample, RNA obtained with NucleoSpin RNA Stool provided better performance in qPCR compared to RNA prepared with the competitor kits.

Overview

Speedy isolation of total RNA from various stool specimens
Combination of mechanical homogenization and chemical lysis ensures optimized RNA yield, including small RNA
Isolated RNA does not show any PCR inhibition, due to NucleoSpin Inhibitor Removal Columns
Suitable for various human and animal stool samples

More Information

Technology	Silica membrane technology combined with NucleoSpin Bead Tubes Type A
Format	Mini spin columns
Starting material	Fresh or frozen stool samples
Fragment size	≥18 nt
Typical yield	10–30 µg
A_260/280	1.9–2.1
A_260/230	1.8–2.5
Typical RIN (RNA integrity number)	>7.5
Elution volume	100 µl
Preparation time	70 min/10 preps
Binding capacity	200 µg

Applications

RNA isolation from stool samples
Typical downstream applications: real-time RT-PCR, northern blotting, primer extension, array technology, RNase protection assays

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

Purify DNA from small samples like microdissected tissue, small blood samples, or small amounts of cells.

Designed for extra small samples

Purify DNA from small samples such as microdissected tissue, small blood samples, or small amounts of cells. Based on reliable silica-membrane technology, NucleoSpin Tissue XS overcomes the limitations of conventional kits using an innovative new column design with a funnel-shaped ring.

Learn more All genomic DNA kits

Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2018 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.