Ultra Deep Microbiome Prep Kit
Ultra-Deep Microbiome Prep isolates bacterial and fungal DNA from body fluids up to 1 ml and from tissue pieces of about 0.5 by 0.5 cm, after depleting at least 95 per cent of the host DNA. Research use only. Because the human DNA is removed before the microbial DNA is purified, the eluate can go into 16S or ITS sequencing and into broad-range PCR without the host background that otherwise dominates the read count.
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This kit in detail
- Removal of at least 95 per cent of host DNA before microbial DNA isolation
- Fluid samples of 0.2 to 1 ml and tissue pieces of about 0.5 by 0.5 cm
- Broad lysis: over 200 bacterial genera and 65 fungal genera
- MolYsis chemistry with MolDNase B and BugLysis
- Spin-column purification, no controls supplied
- 25 or 50 extractions per kit, REF G-020-025 and G-020-050
Ultra-Deep Microbiome Prep isolates bacterial and fungal DNA from clinical material in which human DNA is present in large excess. Removing the host DNA first is what makes low-abundance microbial DNA visible.
Principle
A chaotropic buffer lyses the human cells in the sample while microbial cells stay intact. The human DNA released, including DNA from dead cells, is degraded by the chaotrope-resistant DNase MolDNase B. The microbial cells are then collected and their cell walls degraded with BugLysis, followed by Proteinase K digestion and spin-column purification of the microbial DNA. The manufacturer states removal of at least 95 per cent of host DNA. The lysis step has been shown to work on over 200 bacterial genera, 86 Gram-positive and 120 Gram-negative, and on 65 fungal genera.
Sample material
Body fluids: ascites, bronchoalveolar lavage, blood, cerebrospinal fluid, pleural fluid, pus and synovial fluid, in volumes of 0.2 to 1 ml. Tissue: abscess material, biopsies, heart valves, prostheses and stents, in pieces of about 0.5 by 0.5 cm. For samples of 1 to 10 ml the manufacturer offers Ultra-Deep Microbiome Prep10.
What is and is not supplied
The kit is delivered in two parts: buffers and consumables at room temperature, and the enzymes MolDNase B, BugLysis, Proteinase K and beta-mercaptoethanol frozen. No positive or negative control is supplied; the manual describes how to prepare them, and an internal PCR control is available separately as Ultra-Deep Microbiome Ampli IC (G-130-042). All reagents and consumables are DNA-free. Shelf life is 24 months, and the procedure can be paused after step 5 by freezing.
Regulatory status
This kit is for research use only and not for use in diagnostic procedures.
- Lysis buffer: A solution that helps to break down the cell walls of microorganisms and release DNA.
- Bead tubes: Tubes containing beads that are used to mechanically disrupt the cell membranes of microorganisms.
- Binding buffer: A solution that helps to bind the DNA to a filter membrane.
- Wash buffer: A solution that helps to remove contaminants from the DNA.
- Elution buffer: A solution that helps to release the purified DNA from the filter membrane.
- Spin columns: Columns that are used to filter and purify the DNA.
- Collection tubes: Tubes used to collect the purified DNA.
- Instruction manual: A guide to assist laboratory technicians and scientists in using the kit effectively.
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Instructions for Use
Frequently paired with this kit
Other assays and instruments lab teams add alongside the Ultra Deep Microbiome Prep Kit.
Add this kit to your quote
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