Host DNA contamination impedes the molecular analyses of microbiomes in host samples, such as bodily fluids and swabs. The SPINeasy Host Depletion Microbial DNA Kit provides an easy-to-use workflow to isolate microbial DNA from samples containing high amounts of host DNA. This background reduction of host DNA is achieved through selective lysis of host cells with our specially formulated Host Lysis Buffer. Host DNA, which has been released in the solution, is then removed from the supernatant through centrifugation. A further host depletion treatment provides enzymatic degradation of any remaining host DNA, as well as DNA from dead microbial cells. Finally, the remaining intact cells are lysed through a combination of specially formulated buffers and highly efficient homogenization using FastPrep instruments from MP Biomedicals. Microbial DNA is purified using a convenient silica-membrane spin-column technology workflow and ready for downstream molecular applications.
Kit performance
The following are PCR results of DNA extraction from saliva samples using SPINeasy Host Depletion Microbial DNA Kit, showing effective host DNA depletion and microbial DNA recovery. Both host DNA depletion and bacteria DNA recovery are estimated to be higher than 90% for these samples, using qPCR analyses.
Figure 1: PCR of DNA extracted from saliva samples using SPINeasy Host Depletion Microbial DNA Kit. a) PCR detection of host DNA using human β-globin primers. b) PCR detection of bacterial DNA using 16S primers. M: DNA marker; Lane 1, 3, 5: Total DNA extracted without performing host depletion steps; Lane 2, 4, 6: Host depleted (H. dep) DNA extracted using SPINeasy Host Depletion Microbial DNA Kit; Lane 7: PCR negative control.
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Application Notes
SPINeasy Host Depletion Microbial DNA Kit is designed to overcome the challenges that researchers face when working with heavily contaminated host DNA samples.
Key Applications
Microbiome analysis, Shot-gun metagenomics sequencing, NGS, Genomics, Molecular Biology
| SKU | 116545050 |
| Alternate Names | SPINeasy Host Depletion Microbial DNA Kit; Host DNA Depletion Kit |
| Application Notes | SPINeasy Host Depletion Microbial DNA Kit is designed to overcome the challenges that researchers face when working with heavily contaminated host DNA samples. |
| Base Catalog Number | 65450 |
| Product Line | SPINeasy |
| Components | 12 mL Equilibration buffer,50x Lysing Matrix E, 60 mL Host Lysis Buffer, 6 mL Host Depletion Buffer, 1 Vial Host Depletion Enzyme HDE, 200 µL HDE Reconstitution Buffer, 12 mL Microbial Selection Buffer, 40 mL Microbial Lysis Buffer, 30 mL Wash Buffer HD1, 6 mL Wash Buffer HD2, 6 mL Wash Buffer HD, 50x Column HD with collection tubes |
| Pack Size | 50 Preps |
| Sample Type | Saliva, Swabs, Bodily Fluid |
| Storage | Store at Room Temperature (15-25℃) in a dry environment |
