Stool DNA Isolation Kit: High-Yield, Inhibitor-Free Microbiome DNA
The Stool DNA Isolation Kit (Cat. 27600) provides a rapid and highly reliable method for the extraction of high-quality genomic DNA from fresh or preserved stool samples. Stool samples are notoriously challenging to process due to the high concentration of complex PCR inhibitors, such as humic acids, bile salts, and complex polysaccharides. This kit is specifically designed to overcome these obstacles, ensuring the extraction of pure DNA that is immediately ready for sensitive downstream applications, including real-time PCR, 16S rRNA sequencing, and shotgun metagenomics.
Utilising a proprietary resin matrix technology, the purification process efficiently captures nucleic acids while completely removing contaminating proteins, lipids, and PCR inhibitors. The procedure bypasses the need for hazardous phenol/chloroform extractions or time-consuming alcohol precipitations. Instead, the lysate is passed through a spin column where the DNA selectively binds to the resin, and impurities are washed away. The resulting purified stool DNA is highly concentrated and possesses excellent A260/280 and A260/230 ratios, accurately representing the full diversity of the faecal microbiome without bias.
Key Benefits:
- Inhibitor-Free DNA: Complete removal of PCR inhibitors. Effectively eliminates humic acids and other complex inhibitors commonly found in faecal matter, ensuring robust amplification in downstream assays.
- High Yield and Purity: Maximise your DNA recovery. Consistently delivers higher yields of pure genomic DNA compared to competitor kits, accurately reflecting the microbial community.
- Versatile Sample Input: Process fresh or preserved samples. Fully compatible with fresh stool or samples preserved in Stool Nucleic Acid Collection and Preservation Tubes.
- Rapid and Safe Workflow: No toxic chemicals required. The spin-column format provides a fast extraction process without the use of phenol or chloroform.
- Broad Downstream Compatibility: Ready for advanced analysis. Purified DNA is ideal for 16S rRNA gene sequencing, metagenomics, PCR, qPCR, and Southern blotting.
Technical Insights
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Figure 1
Higher Yields of DNA than Competitor Q. Stool DNA was isolated from 200 mg of fresh or preserved stool samples using Norgen’s Stool DNA Isolation Kit and Competitor Q’s Kit. For evaluation, 10 µL of DNA from the elution was run on 1X TAE 1.2% agarose gel. Norgen’s kit isolated much higher yields of DNA. *Stool was collected using Norgen’s Stool Nucleic Acid Collection and Preservation tubes (Cat. 45660). Marker = Norgen’s HighRanger DNA Ladder (Cat. 11900).
Figure 2
Stool DNA quality and concentration measured by NanoDrop. High DNA concentration and quality was obtained using Norgen’s Stool DNA Isolation Kit from fresh or preserved stool samples. Figure 2A: Fresh stool samples, Figure 2B: Preserved stool using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660).
Figure 3
Detection of 16S rRNA from stool DNA isolated from 200 mg of fresh stool samples using Norgen’s Stool DNA Isolation Kit and Competitor Q’s kit. DNA quality was confirmed by Real-time PCR using 2 µL of stool DNA (total PCR reaction volume was 20 µL) to detect 16S rRNA from different microorganisms in the stool samples. The earlier Ct value with Norgen’s DNA samples (blue lines) compared to Competitor Q’s samples (red lines) indicated a higher quality of stool DNA for downstream applications. Figure 3A: Fresh stool sample, Figure 3B: Preserved stool using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660).
Figure 4
Detection of 5S rRNA from the Stool DNA isolated from 200 mg of fresh stool samples using Norgen’s Stool DNA Isolation Kit and Competitor Q’s Kit. DNA quality was confirmed by Real-time PCR using 2 µL of stool DNA (total PCR reaction volume was 20 µL) to detect 5S rRNA from eukaryotic DNA in the stool samples. The earlier Ct value with Norgen’s DNA samples (blue lines) compared to Competitor Q’s samples (red lines) indicated a higher quality of stool DNA for downstream applications. Figure A: Fresh stool sample, Figure B: Preserved stool using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660).
Figure 5
Better 16S rRNA detection from DNA isolated using Norgen’s Stool DNA Preservative and Stool DNA Isolation Kit. Two microlitres of stool DNA was used in a 20 µL PCR reaction volume. Stool was preserved using Norgen’s Stool DNA Preservative and isolated using Norgen’s Stool DNA Isolation Kit showed better 16S rRNA gene detection as compared to fresh stool isolated with Competitor Q.
Figure 6
Distribution of 10 Different Fecal Microbiomes. A) Principal Coordinate Analysis of 10 fecal microbiomes showing differences in the distribution of taxonomic classifications between samples up to class level. B) Hierarchical clustering of 10 fecal microbiomes based on genus-level classifications, including a bar chart showing the relative abundance of genus-level classifications for each sample in the dendrogram.
Figure 7
High Quality DNA Isolated from Preserved Stool Samples. DNA was isolated from 200 µL preserved stool samples using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System). For evaluation, 10 µL from 75 µL of elution were run on 1X TAE 1.2% agarose gel. Stool samples were preserved in Norgen’s Stool Nucleic Acid Collection and Transport Tubes (Cat. 45630, 45660). M = Norgen’s HighRanger DNA Ladder (Cat. 11900).
Figure 8
Comparison of A260/280 and A260/230 ratios. DNA was isolated from 200 mg stool samples using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System) and Norgen’s Stool DNA Isolation Kit (column format). The purified DNA was then compared for A260/280 and A260/230 ratios. DNA isolated using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System) (A, B and C) showed a comparable DNA quality to the DNA isolated using Norgen’s Stool DNA Isolation (Column method), indicating the consistence and highest DNA quality were generated by the Stool DNA Isolation Kit (Magnetic Bead System).
Figure 9
High Quality DNA confirmed by Real-time PCR. DNA was isolated from 200 mg stool using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System) and Norgen’s Stool DNA Isolation Kit (column format). The quality of the purified DNA was evaluated by using 8 μL of stool DNA (total PCR reaction volume was 20 µL) to detect 16S rRNA from the different stool samples. No PCR inhibition was observed from DNA isolated using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System) (blue circles), similar to the DNA isolated using Norgen’s Stool DNA Isolation Kit (Column method) (red circles), indicating the excellent isolation consistency and the quality of the stool DNA for downstream applications.
Figure 10
16S Metagenomics data generated by Illumina MiSeq. Stool DNA was isolated using Norgen’s Stool DNA Isolation Kit (Magnetic Bead System) from 200 mg of stool from healthy donors, and the stool microbiomes were successfully sequenced by Illumina MiSeq.
Figure 11
High Quality DNA Isolated from Preserved Stool Samples. DNA was isolated from 200 µL preserved stool samples using Norgen’s Stool DNA Isolation 96-Well Kit (Magnetic Bead System). For evaluation, 10 µL from 75 µL of elution were run on 1X TAE 1.2% agarose gel. M = Norgen’s HighRanger DNA Ladder (Cat. 11900).
Figure 12
High Quality DNA Confirmed by Real-Time PCR. DNA was isolated from 200 mg stool using Norgen’s Stool DNA Isolation 96-Well Kit (Magnetic Bead System). The quality of the purified DNA was evaluated by using 2 µL of stool DNA (total PCR reaction volume was 20 µL) to detect 16S rRNA from the stool sample. No PCR inhibition was observed from DNA isolated using Norgen’s Stool DNA Isolation 96-Well Kit (Magnetic Bead System) indicating the excellent isolation consistency and the high quality of the stool DNA for downstream applications.
Figure 13
Comparison of DNA yield and integrity using two automated platforms with the Stool DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System, Cat. 63100). DNA was extracted from 200 µL preserved stool samples using the same extraction kit on two automated platforms: Isopure 96 and KingFisher. Total DNA yield was assessed by 16S rRNA gene qPCR (left panel), showing comparable performance across platforms and replicates. Error bars represent standard deviations. High-molecular-weight DNA was consistently recovered from both platforms, demonstrating effective and reproducible DNA isolation regardless of automation system used.
Specifications
- Sample Input: Up to 200 mg of fresh/frozen stool or 200 µL of preserved stool
- Elution Volume: 50 µL
- Processing Time: 30 minutes
- Technology: Spin column chromatography
- DNA Quality: Free from PCR inhibitors (humic acid, etc.)
- Storage Conditions: Room temperature