Microbiome analysis involves studying the composition, diversity, and functional potential of microbial communities that inhabit various environments, including the human body, soil, water, and other ecosystems.
The human microbiome, in particular, refers to the collection of microorganisms (bacteria, viruses, fungi, and more) that live on and inside the human body, influencing various aspects of health and disease.
Microbiome analysis is conducted to understand the complex interactions between these microbes and their host, and how these interactions impact health, disease, and ecosystem functioning.
The MagSi-DNA Stool kit is intended for manual and automated isolation of microbiome DNA (mixture of bacterial/microbial and host DNA) from fresh, frozen or stabilized human stool samples.
Procedure
Processing time for the preparation of 96 samples is about 60 minutes including heat lysis incubations. The obtained DNA can be used directly for downstream applications such as PCR or NGS.
MagSi-DNA Stool is optimized for use in isolating total DNA from human stool samples. The MagSi-ST10 magnetic beads are easy to handle and are supplied in a storage buffer for optimized resuspension and minimized sedimentation. Depending on the sample materials RNA may be co-purified.
For efficient lysis of hard-to-lyse bacteria (e.g. gram positive bacteria) mechanical disruption with GP Lysis Tubes is recommended. If the microorganism of interest requires stronger homogenization than provided by a vortex, or if using a bead beater is desired, highpowered bead beating may be used, e.g. with the Geno/Grinder® (SPEX Sample Prep)
Easy to automate
Figure 1. Nucleic acid concentrations (NanoDrop) obtained from different input weights of stool samples. A260/280 ratios are 1.95 for all sample input weights (data not shown).
Figure 2. Nucleic acid concentrations (NanoDrop) obtained from stool stabilized in OMNIgene®•GUT. 200 µL of the preserved stool sample was processed using MagSi-DNA Stool versus two competitor kits.
Figure 3. Ct values obtained by qPCR targeting a bacterial 16s rDNA gene on purified DNA obtained from preserved stool samples versus competitor extraction kits. Purified samples were used undiluted and diluted as indicated in the qPCR reaction. No PCR inhibition was observed. Lowest Ct values were obtained with the MagSi-DNA Stool kit, demonstrating the high yield of the recovered DNA.
Figure 4. Electropherogram (TapeStation) obtained from stool stabilized in OMNIgene®•GUT, showing recovery of high-molecular weight DNA. 200 µL of the preserved stool sample was processed using MagSi-DNA Stool.
ART.NO. | DESCRIPTION | AMOUNT |
|---|---|---|
MDKT00230096 | MagSi-DNA Stool | 96 preps |
MDKT00230960 | MagSi-DNA Stool | 10 x 96 preps |
MDKT0023B096 | MagSi-DNA Stool (incl. GP Lysis Tubes) | 96 preps |
MDKT0023B960 | MagSi-DNA Stool (incl. GP Lysis Tubes) | 10 x 96 preps |
MDPL00330100 | GP Lysis Tubes | 100 pcs |