DNeasy PowerSoil Pro Kit (50)，47014，Qiagen，凯杰

DNeasy PowerSoil Pro Kit

For the isolation of microbial genomic DNA from all soil types

Efficient lysis of bacteria and fungi in all soil types

Up to 8-fold higher DNA yields compared to alternative methods

Recovery of inhibitor-free DNA for direct use in NGS applications

Unbiased results accurately represent sample alpha diversity

Automation on the QIAcube

Figure 1: Isolate more high-quality genomic DNA.

250 mg of the three different soil types were prepared using commercially available sample preparation solutions and compared to the new DNeasy PowerSoil Pro Kit. The DNeasy PowerSoil Pro Kit out-performed other available kits, resulting in 8-fold greater recovery of DNA across the variety of soil types.

Figure 3: Increased bacterial OTUs with the new DNeasy PowerSoil Pro Kit.

DNA prepared from soil samples was isolated with different methods and enriched. Analysis of the 16S rRNA-genes was done using the QIAseq 1-Step Amplicon Kit and data analysis was done using the Microbial Genomics Pro suite (CLC workbench). Alpha diversity was determined by total number of bacterial operational taxonomic units (OTUs). The greatest diversity was seen in the samples processed with the DNeasy PowerSoil Pro Kit.

Figure 2: Improved, high-quality DNA yields with the new DNeasy PowerSoil Pro Kit.

Gel A shows 1.8 ml of Candida albicans fungal culture processed with the DNeasy PowerSoil Kit (a) compared with the DNeasy PowerSoil Pro Kit (b). Gels B and C contain various soil samples (250 mg). Gel B was processed with the original DNeasy PowerSoil Kit, while Gel C was processed using the DNeasy PowerSoil Pro Kit. As evidenced by the stronger gel bands, a greater DNA yield was isolated from the sample using the DNeasy PowerSoil Pro Kit.

Figure 4: Increased purity and less variability in DNA extracted with the DNeasy PowerSoil Pro Kit.

Soil samples (250 mg each) were prepared using commercially available sample preparation solutions and compared to the new DNeasy PowerSoil Pro Kit. DNA purity was depicted via UV measurements and the analysis of ratios A260/A280, A, and A260/A230, B. Inhibitor removal was visualized with an inhibitor-sensitive PCR with an internal control spiked with eluates from the PureLink Microbiome DNA Purification Kit and the DNeasy PowerSoil Pro Kit. Eluate-free reactions served as internal control, C. DNA extracted with the DNeasy PowerSoil Pro Kit shows an increased purity compared with other sample preparation kits with no inhibition of PCR.

Extracting microbial DNA from soil samples can be challenging. QIAGEN’s new DNeasy PowerSoil Pro Kit is even more effective than our original PowerSoil technology at isolating high yields of pure microbial DNA from all soil types, including compost, clay and top soil. The kit features a novel bead tube and optimized chemistry for more efficient lysis of soil bacteria and fungi. The kit also contains streamlined Inhibitor Removal Technology (IRT) to eliminate the challenging inhibitors commonly found in soil and environmental samples in even less time. Sequencing results reveal higher alpha diversity as measured by observed operational taxonomic units (OTUs) compared to other tested methods. Extraction of DNA using the DNeasy PowerSoil Pro Kit can be automated on the QIAcube Connect.

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Performance

Significantly increased DNA yields from challenging samples

The DNeasy PowerSoil Pro Kit includes a novel bead tube and improved lysis chemistry, which enables isolation of up to 8-fold higher yields of DNA compared to the first generation DNeasy PowerSoil Kit and those of competitors in all soil types tested (See: ' Isolate more high-quality genomic DNA.'). Both the DNeasy PowerSoil Kit and the new DNeasy PowerSoil Pro Kit isolated high-quality DNA from challenging soil types (See: ' Improved, high-quality DNA yields with the new DNeasy PowerSoil Pro Kit.').

Unbiased identification of total diversity and community representation in soil samples

DNA isolated using the DNeasy PowerSoil Pro Kit identified more bacteria (OTUs) in soil samples when compared with the DNeasy PowerSoil Kit and those of two competitors (See: ' Increased bacterial OTUs with the new DNeasy PowerSoil Pro Kit.').

Principle

Inhibitor Removal Technology increases purity of isolated DNA

The DNeasy PowerSoil Pro Kit features a streamlined Inhibitor Removal Technology (IRT) to decrease sample processing time. In a comparison of methods for DNA isolation from soil, the DNeasy PowerSoil Pro Kit was the only method showing 260/280 ratios near 1.8 for all soil types, as well as the highest 260/230 ratios, indicating the absence of inhibitors. Samples isolated using the kit also showed no inhibition in PCR, in contrast to competitor methods (See: ' Increased purity and less variability in DNA extracted with the DNeasy PowerSoil Pro Kit.').

Procedure

The soil or stool samples are lysed via chemical and mechanical homogenization. Lysis buffer is added to a mixed zirconium bead tube containing the sample. Bead beating can be carried out using a standard benchtop vortex with bead tube adapter or the high-powered PowerLyser. Crude lysate is then subjected to inhibitor removal for cleanup. Following the cleanup, purified lysate is mixed with an equal volume of DNA binding solution and passed through a silica spin filter membrane. The membrane is washed with a two-step washing regime. Silica-bound DNA is then eluted using a 10 mM Tris elution buffer. Relevant downstream applications for the isolated DNA include PCR, NGS and enzymatic digestion assays.