粪便 DNA 分离试剂盒

概述

这些试剂盒提供了一种方便快捷的方法，可从新鲜、冷冻和保存的粪便样本（包括使用 Norgen 的粪便核酸采集和保存剂试管保存的样本）中分离出总 DNA。45660）。该通用方案可同时从粪便样本中发现的各种微生物和宿主细胞中分离出总基因组 DNA，非常方便。纯化 DNA 的产量高，分子量可达 50 kb 以上。纯化后的 DNA 可用于多种下游应用。

粪便 DNA 分离试剂盒（旋转柱）

同时检测粪便样本中微生物和宿主细胞 DNA 的通用方法。通过化学和物理均质与裂解相结合的方法，消除聚合酶链式反应抑制剂，包括所有腐植酸痕迹。然后使用简单快速的旋转柱程序进一步纯化 DNA。由于在分离过程中去除了所有腐殖酸物质和其他聚合酶链式反应抑制剂，因此纯化后的 DNA 质量最高，与所有下游应用（如 PCR、qPCR、NGS 和微阵列）完全兼容。

粪便 DNA 分离 96 孔试剂盒（高通量）

使用离心法进行快速、简便的高通量处理。通过化学和物理均质与裂解相结合的方法，消除聚合酶链式反应抑制剂，包括所有腐植酸痕迹。然后使用简单快速的旋转柱程序进一步纯化 DNA。由于在分离过程中去除了所有腐殖酸物质和其他聚合酶链式反应抑制剂，因此纯化后的 DNA 质量最高，与所有下游应用（如 PCR、qPCR、NGS 和微阵列）完全兼容。

粪便 DNA 分离试剂盒（磁珠系统）

使用磁珠系统进行快速简便的加工。强大的裂解系统（化学裂解与机械均质相结合）。

粪便 DNA 分离 96 孔试剂盒（高通量磁珠系统）

该试剂盒使用磁珠系统和强大的裂解系统（化学裂解与机械均质相结合）进行快速简便的处理。吞吐量高，与自动化机器人系统兼容。

细节

Supporting Data

Figure 1 / 13

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 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.

支持数据

图 1.使用 Norgens Stool DNA Isolation Kit 和竞争对手 Q 的试剂盒从 200 mg新鲜或保存的粪便样本中分离出 DNA。为了进行评估，将洗脱出的 10 µL DNA 在 1X TAE 1.2% 琼脂糖凝胶上运行。Norgen 试剂盒分离出的 DNA 产量要高得多。*使用 Norgens 粪便核酸采集和保存剂试管（类别号：45660）。标记 = Norgens HighRanger DNA Ladder（类别号：11900）。

图 2.用 NanoDrop 测量粪便 DNA 质量和浓度。使用 Norgens 粪便 DNA 分离试剂盒从新鲜或保存的粪便样本中获得高浓度和优质 DNA。图 2A：新鲜粪便样本，图 2B：使用 Norgens 粪便核酸采集和保存剂试管（类别号：45660）。

图 3.使用 Norgen 的粪便 DNA 分离试剂盒和竞争对手 Qs 试剂盒，从 200 mg新鲜粪便样本中分离出的粪便 DNA 中检测 16S rRNA。使用 2 µL 粪便 DNA（聚合酶链式反应总体积为 20 µL）进行实时聚合酶链式反应，以检测粪便样本中不同微生物的 16S rRNA，从而确认 DNA 质量。与竞争对手的 Qs 样本（红线）相比，Norgen 的 DNA 样本（蓝线）的 Ct 值更高，这表明用于下游应用的粪便 DNA 质量更高。图 3A：新鲜粪便样本，图 3B：使用 Norgens 粪便核酸采集和保存剂试管（类别号：45660）。

图 4.使用 Norgen 的粪便 DNA 分离试剂盒和竞争对手 Qs 试剂盒从 200 mg新鲜粪便样本中分离出的粪便 DNA 中检测 5S rRNA。使用 2 µL 粪便 DNA（聚合酶链式反应总体积为 20 µL）进行实时聚合酶链式反应，检测粪便样本中真核 DNA 的 5S rRNA，从而确认 DNA 的质量。与竞争对手的 Qs 样本（红线）相比，Norgen 的 DNA 样本（蓝线）的 Ct 值更高，这表明用于下游应用的粪便 DNA 质量更高。图 4A：新鲜粪便样本，图 4B：使用 Norgens 粪便核酸采集和保存剂试管（类别号：45660）。

图 5.使用 Norgens 粪便 DNA 保存剂和粪便 DNA 分离试剂盒从 DNA 中分离出的 16S rRNA 检测效果更佳。在 20 µL 聚合酶链式反应体积中使用了 2µL 粪便 DNA。与使用竞争对手 Q 分离的新鲜粪便相比，使用 Norgens 粪便 DNA 保存剂保存并使用 Norgens 粪便 DNA 分离试剂盒分离的粪便显示出更好的 16S rRNA 基因检测结果。

图 6.10 种不同粪便微生物群的分布。A) 对 10 个粪便微生物组进行的主坐标分析显示，不同样本之间的分类学分类分布存在差异，最高可达类别级别。B) 根据属级分类对 10 个粪便微生物组进行分级聚类，包括显示树枝图中每个样本属级分类相对丰度的柱状图。

图 7.从保存的粪便样本中分离出优质 DNA。使用 Norgens 粪便 DNA 分离试剂盒（磁珠系统）从 200 µL 保存的粪便样本中分离出 DNA，并将 75 µL 洗脱液中的 10 µL 在 1X TAE 1.2% 琼脂糖凝胶上进行检测。粪便样本保存在 Norgens 粪便核酸采集和运输管（Cat.45630、45660）。M = Norgens HighRanger DNA Ladder（商品编号：Cat.11900）。

图 8.A260/280 和 A260/230 比率的比较。使用 Norgen 粪便 DNA 分离试剂盒（磁珠系统）和 Norgen 粪便 DNA 分离试剂盒（柱式）从 200 mg粪便样本中分离 DNA。然后比较纯化 DNA 的 A260/280 和 A260/230 比率。使用诺健斯粪便 DNA 分离试剂盒（磁珠系统）分离的 DNA（A、B 和 C）与使用诺健斯粪便 DNA 分离试剂盒（柱法）分离的 DNA 质量相当，表明粪便 DNA 分离试剂盒（磁珠系统）产生的 DNA 具有一致性和最高的 DNA 质量。

图 9.经实时聚合酶链式反应验证的优质 DNA。使用 Norgen 粪便 DNA 分离试剂盒（磁珠系统）和 Norgen 粪便 DNA 分离试剂盒（柱式）从 200 mg粪便中分离 DNA。使用 8 升粪便 DNA（聚合酶链式反应总体积为 20 µL）检测不同粪便样本中的 16S rRNA，以评估纯化 DNA 的质量。使用诺健粪便 DNA 分离试剂盒（磁珠系统）分离出的 DNA（蓝色圆圈）与使用诺健粪便 DNA 分离试剂盒（柱法）分离出的 DNA（红色圆圈）相似，未发现聚合酶链式反应抑制现象，表明分离一致性极佳，可为下游应用提供优质粪便 DNA。

图 10.由 Illumina MiSeq 生成的 16S 元基因组学数据。使用 Norgens 粪便 DNA 分离试剂盒（磁珠系统）从健康供体的 200 mg粪便中分离出粪便 DNA，并通过 Illumina MiSeq 成功对粪便微生物组进行测序。

图 11.从保存的粪便样本中分离出优质 DNA。使用 Norgens 粪便 DNA 分离 96 孔试剂盒（磁珠系统）从 200 µL 保存的粪便样本中分离 DNA。在 1X TAE 1.2% 琼脂糖凝胶上从 75 µL 洗脱液中提取 10 µL 进行评估。M = Norgens HighRanger DNA Ladder（商品编号：Cat.11900）。

图 12.通过实时聚合酶链式反应确认优质 DNA。使用 Norgen 的粪便 DNA 分离 96 孔试剂盒（磁珠系统）从 200 mg粪便中分离 DNA。使用 2 µL 粪便 DNA（聚合酶链式反应总体积为 20 µL）检测粪便样本中的 16S rRNA，以评估纯化 DNA 的质量。使用 Norgen 的粪便 DNA 分离 96 孔试剂盒（磁珠系统）分离出的 DNA 未出现聚合酶链式反应抑制现象，这表明该试剂盒的分离一致性极佳，可为下游应用提供优质粪便 DNA。

试剂盒规格
最粪便便输入量	200 mg（新鲜/冷冻粪便）或 400 μL （保存粪便
粪便加工类型	冷冻、新鲜或保存粪便
形式	旋转栏
色谱柱最大结合能力	50 μg
最大色谱柱装载量	650 μL
洗脱体积	50 μL
完成 10 次净化所需的时间	30 分钟
应用	PCR、qPCR、Southern 印迹分析、测序、微阵列分析。

储存条件和产品稳定性
所有溶液都应密封保存在室温下。该试剂盒自发货之日起 2 年内保持稳定。

Documentation

Alternate formats available upon request by emailing info@norgenbiotek.com. Such requests shall be made:

in a timely manner that considers the person's accessibility needs due to disability; and
at a cost that is no more than the regular cost charged to other persons

概述

细节

Supporting Data

Figure 1. Higher Yields of DNA than Competitor Q.

Figure 2. Stool DNA quality and concentration measured by NanoDrop.

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.

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.

Figure 5. Better 16S rRNA detection from DNA isolated using Norgen’s Stool DNA Preservative and Stool DNA Isolation Kit.

Figure 6. Distribution of 10 Different Fecal Microbiomes.

Figure 7. High Quality DNA Isolated from Preserved Stool Samples.

Figure 8. Comparison of A260/280 and A260/230 ratios.

Figure 9. High Quality DNA confirmed by Real-time PCR.

Figure 10. 16S Metagenomics data generated by Illumina MiSeq.

Figure 11. High Quality DNA Isolated from Preserved Stool Samples.

Figure 12. High Quality DNA Confirmed by Real-Time PCR.

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).

支持数据

Documentation

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