Bacterial and host DNA isolation from stool samples

Norgen's Stool DNA Isolation Kits offer quick and easy means for the purification of bacterial and host DNA from stool and fecal samples.

Format

Spin column

Size

50 preps

Image Credit: Norgen Biotek Corp.

Image Credit: Norgen Biotek Corp.

High throughput

Size

2 × 96-Well Plates

Image Credit: Norgen Biotek Corp.

Magnetic bead system

Size

50 preps

Image Credit: Norgen Biotek Corp.

Image Credit: Norgen Biotek Corp.

High throughput magnetic bead system

Size

2 × 96-Well Plates

Image Credit: Norgen Biotek Corp.

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Features and benefits

• Simultaneous isolation of DNA from both the host and microbes using a universal technique
• Compatible with stool samples that are fresh, frozen, and/or preserved
• High-quality DNA: Get HMW DNA up to 50 kb and above. Suitable for PCR, qPCR, sequencing, and microarray applications.
• Eliminates humic acids and other PCR inhibitors
• Integrates chemical and physical homogenization to ensure efficient sample lysis
• Available in several formats to suit your preferred workflow

Available formats. Source: Norgen Biotek Corp.

Details

Supporting data

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). Image Credit: Norgen Biotek Corp.

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). Image Credit: Norgen Biotek Corp.

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). Image Credit: Norgen Biotek Corp.

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 4A: Fresh stool sample, Figure 4B: Preserved stool using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660). Image Credit: Norgen Biotek Corp.

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. Image Credit: Norgen Biotek Corp.

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. Image Credit: Norgen Biotek Corp.

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). Image Credit: Norgen Biotek Corp.

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). Image Credit: Norgen Biotek Corp.

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. Image Credit: Norgen Biotek Corp.

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. Image Credit: Norgen Biotek Corp.

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). Image Credit: Norgen Biotek Corp.

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. Image Credit: Norgen Biotek Corp.

Automation

Stool DNA Automated Isolation tutorial

Video Credit: Norgen Biotek Corp.

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Spin column

Source: Norgen Biotek Corp.

Storage Conditions and Product Stability: All solutions should be stored at room temperature and kept tightly sealed. This kit is stable for two years from the dispatch date.

High throughput

Source: Norgen Biotek Corp.

Storage Conditions and Product Stability: All solutions must be stored at room temperature and kept tightly sealed. This kit is stable for one year from the dispatch date.

Magnetic bead system

Source: Norgen Biotek Corp.

* Average DNA yield will vary depending on the donor

Storage Conditions and Product Stability: All solutions need to be stored at room temperature and kept tightly sealed. This kit is stable for one year following the date of shipment.

High throughput magnetic bead system

Source: Norgen Biotek Corp.

* Average DNA yield will vary depending on the donor

Storage Conditions and Product Stability: All solutions need to be stored at room temperature and kept tightly sealed. This kit is stable for one year from the dispatch date.

Source: Norgen Biotek Corp.