Introduction
MiniGenomics® Stool DNA Isolation Kit is designed for rapid extraction and purification of high-quality, ready-to-use genomic DNA from fresh or frozen stool samples.
Features
o Advanced magnetic nanoparticle technology for superior DNA adsorption and release
o Standard purification methods suitable for biological samples in various forms or reagents
o Specialized reagents to enhance DNA quality from challenging samples
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Compatible with multiple high-throughput automated nucleic acid extraction platforms
Performance of Isolating DNA from Stool Samples
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Exhibit 1: DNA extraction of stool samples from 4 volunteers by MiniGenomics® Stool DNA Isolation Kit and a competitor kit (Q brand). DNA product (1/20) was detected by agarose gel electrophoresis. |
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Exhibit 2: qPCR detection of human β-globin gene level in extracted DNA of all samples. PCR product (1%) were analyzed by agarose gel electrophoresis. Ct values are shown in the table below. |
Table 1: Qualitative and quantitative analysis of gDNA extracted from the stool samples on Nanodrop 2000. |
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Performance of Isolating Pathogen DNA from Stool Samples
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Exhibit 3: DNA extraction of stool samples from 2 volunteers by MiniGenomics® Stool DNA Isolation Kit. DNA product (1/20) was detected by agarose gel electrophoresis. |
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Exhibit 4: qPCR detection of bacterial 16SrRNA gene level in extracted DNA of all samples. PCR product (1%) were analyzed by agarose gel electrophoresis. |
Products
Sample type
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Fresh or frozen stool sample |
Cat #
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A181206-50 (50 preps)
A181206-200 (200 preps)
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Capacity
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50 preps or 200 preps
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Manual protocol
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Protocol download
(A181206-50 for 50 preps)
Protocol download
(A181206-200 for 200 preps)
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Time per run (manual)
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In total 50~70 minutes for manual operation, including - 20~40 minutes pre-treatment and lysis time depending on sample size and status - 10 minutes binding time - 15 minutes washing time - 5 minutes elution time |
MSDS
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Download
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MagTouch 2000 automation programs & instructions
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Automation protocol download
Program Installation instruction:
1. Download "Items.zip" file and unzip, save the folder to the root directory of a flash drive.
2. Plug the flash drive into the USB port at the back of MagTouch 2000.
3. Press Settings > Import & export > Import. Select the program(s) by pressing the checkmark once (check mark symbol will turn green) and press import.
Any questions? Please send an email to support@apostlebio.com
or call support hotline at 888-305-3218 (Mon-Fri: 9AM to 6PM, PST)
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References
- Combined detection of SDC2/ADHFE1/PPP2R5C methylation in stool DNA for colorectal cancer screening. Journal of Cancer Research and Clinical Oncology June 03, 2023.
- A systematic evaluation of stool DNA preparation protocols for colorectal cancer screening via analysis of DNA methylation biomarkers. Clinical Chemistry and Laboratory Medicine 2021; 59(1): 91–99.
- CE marked colon cancer test kits (as OEM supplier)
- More ...
Combined detection of SDC2/ADHFE1/PPP2R5C methylation in stool DNA for colorectal cancer screening. Li B, et al. Journal of Cancer Research and Clinical Oncology. June 03, 2023. https://doi.org/10.1007/s00432-023-04943-4
Background Colorectal cancer (CRC) is a disease of global concern, and its increasing incidence suggests the need for early and accurate diagnosis. The aim of this study was to investigate the value of combined detection of SDC2, ADHFE1 and PPP2R5C gene methylation in stool samples for early CRC screening.
Methods StoolsamplesfrompatientswithCRC(n=105),advancedadenoma(AA)(n=54),non-advancedadenoma(NA) (n = 57), hyperplastic or other polyps (HOP) (n = 47) or no evidence of disease (NED) (n = 100) were collected from Sep- tember 2021 to September 2022. The methylation levels of SDC2, ADHFE1 and PPP2R5C were quantified by quantitative methylation-specific polymerase chain reaction (qMSP), and faecal immunochemical testing (FIT) was performed. The diagnostic value was assessed using reporter operating characteristic (ROC) curve analysis.
Results The sensitivity of combined detection of SDC2/ADHFE1/PPP2R5C methylation in predicting CRC (0–IV) was 84.8%, the specificity was 98.0%, and the AUC was 0.930 (95% CI 0.889–0.970). Compared to FIT and serum tumour bio- markers, it showed better diagnostic performance for different stages of CRC.
Conclusion The results of this study verified that the methylation levels of SDC2, ADHFE1 and PPP2R5C in stool DNA were significantly increased in CRC patients. Combined detection of SDC2/ADHFE1/PPP2R5C methylation is a potential non-invasive diagnostic method for CRC and precancerous lesion screening.
Clinical trial registration Chinese Clinical Trials Registry, ChiCTR2100046662, registered on 26 May 2021, prospective registration.
(Methods section) DNA isolation, bisulphite treatment and fluorescence‐based quantitative methylation‐specific polymerase chain reaction (qMSP)
Genomic DNA was isolated from stool samples using the Apostle MiniGenomics (TM) Stool Fast Kit (Apostle, A181206) and stored in the refrigerator at – 20 °C until further use.
Jin et al. Clinical Chemistry and Laboratory Medicine (CCLM). 2021; 59(1): 91–99
Objectives - Colorectal cancer (CRC) screening using stool samples is now in routine use where tumor DNA methylation analysis for leading markers such as NDRG4 and SDC2 is an integral part of the test. However, processing stool samples for reproducible and efficient extraction of human genomic DNA remains a bottleneck for further research into better biomarkers and assays.
Methods - We systematically evaluated several factors involved in the processing of stool samples and extraction of DNA. These factors include: stool processing (solid and homogenized samples), preparation of DNA from supernatant and pellets, and DNA extraction with column and magnetic beads-based methods. Furthermore, SDC2 and NDRG4 methylation levels were used to evaluate the clinical performance of the optimal protocol.
Results - The yield of total and human genomic DNA (hgDNA) was not reproducible when solid stool scraping is used, possibly due to sampling variations. More reproducible results were obtained from homogenized stool samples. Magnetic beads-based DNA extraction using the supernatant from the homogenized stool was chosen for further analysis due to better reproducibility, higher hgDNA yield, lower non-hgDNA background, and the potential for automation. With this protocol, a combination of SDC2 and NDRG4 methylation signals with a linear regression model achieved a sensitivity and specificity of 81.82 and 93.75%, respectively.
Conclusions - Through the systematic evaluation of different stool processing and DNA extraction methods, we established a reproducible protocol for analyzing tumor DNA methylation markers in stool samples for colorectal cancer screening.
(Methods section) For magnetic beads-based method, Apostle Stool gDNA Isolation Kit (APOSTLE) was used according to the manufacturer’s instructions. Either 0.2 g pellets or 0.2 mL supernatant from homogenized stool was mixed with 1 mL lysis buffer (APOSTLE) for DNA extractions.
BGI’s Three Complementary Kits of Colorectal Cancer Testing Have Been CE Marked.
BGI. July 13, 2021
BGI Genomics announces that its Stool Sample Collection Kit, DNA Isolation Kit together with Sample Pretreatment Kit for Methylation Detection have been CE marked.
All three kits are used in conjunction with the previously CE marked Colorectal Cancer Testing Product which can detect the methylation of SDC2, ADHFE1 and PPP2R5C genes in human fecal samples.
According to the Global Cancer 2020 (GLOBOCAN) statistics, there are about 19.3 million new cases of colorectal cancer each year, accounting for 10 percent of all new cancer cases. About 935,000 colorectal cancer deaths occur each year, accounting for 9.4 percent of all cancer deaths.
So far, BGI has obtained CE mark for all four products used in the colorectal cancer detection workflow, from sample collection, DNA extraction, DNA pre-treatment to methylation detection, providing customers with reliable and standardized reagents and services.
(Note: Apostle is the Original Equipment Manufacturer or OEM for the Stool DNA Isolation Kit mentioned in this news. Apostle 's branded product is called Apostle MiniGenomics Stool Fast Kit.)