MagAttract PowerSoil Pro DNA Kit

The MagAttract PowerSoil Pro DNA Kit comprises a novel and proprietary method for isolating microbial genomic DNA from environmental and stool samples. The kit uses QIAGEN’s second-generation Inhibitor Removal Technology® (IRT) and is intended for use with environmental samples containing high humic acid content, including difficult soil types such as compost, sediment, and manure. Other more common soil and stool types have also been used successfully with this kit. Improved IRT combined with more efficient bead beating and lysis chemistry yields high-quality DNA that can be used immediately in downstream applications, including PCR, qPCR, and next-generation sequencing (16S and whole-genome).

The MagAttract PowerSoil Pro DNA Kit allows automated high-throughput isolation of DNA from up to 384 soil samples in less than 1 day and is optimized for use with the Thermo Scientific® KingFisher® Flex platform. Protocols for other instruments are being developed.

The MagAttract PowerSoil Pro DNA Kit is effective at removing PCR inhibitors from soil and stool materials from even the most difficult types. Environmental or human samples are added to a 96-well bead beating plate or bead beating tubes for rapid and thorough homogenization. Cell lysis occurs by a combination of mechanical and chemical methods. Humic acids are removed using IRT. Prepared lysates are transferred to the KingFisher Flex platform where total genomic DNA is captured on specialized magnetic beads in the presence of buffers. DNA is washed on the beads and then eluted, and is ready for NGS, PCR, and other downstream applications.

The MagAttract PowerSoil Pro DNA Kit has demonstrably improved performance over legacy kits (See "Performance comparison of MagAttract PowerSoil Pro DNA to the legacy MagAttract PowerSoil and the manual DNeasy PowerSoil Pro Kit") and competitor kits (See "Performance comparison of MagAttract PowerSoil Pro DNA Kit to different competitor kits")

Starting material

This kit is intended primarily for the extraction of microbial genomic DNA from complex substrates, such as soil or stool. Typically, these samples contain high amounts of inhibitors that interfere with downstream enzymatic reactions and compounds that can degrade DNA. The MagAttract PowerSoil Pro DNA Kit is specially designed for the removal of these substances and for the extraction of microbial genomic DNA that is free of proteins, nucleases, and other contaminants or inhibitors (See "DNA quality, as measured by QIAxpert" and "Co-isolation of inhibitors assessed via inhibition PCR").

The recommended starting amount of stool sample is 100 mg. The maximum recommended starting material for stool samples is 200 mg; for dehydrated stool samples, start with no more than 100 mg. In general, highest yields are seen with 50–150 mg of input stool material, though this varies with the source. The recommended starting amount of soil material is 250 mg. Microbial genomic DNA purified using the MagAttract PowerSoil Pro DNA Kit is ready for use in enzymatic reactions, such as PCR or NGS, or it can be stored at −30 to −15°C.

Bead beating

Bead beating is required for efficient lysis of microbial cells. The MagAttract PowerSoil Pro DNA Kit is designed to be used in combination with either PowerBead Pro Plates (cat. no. 19311) or PowerBead Pro Tubes (cat. no. 19301). The use of other disruption media can lead to reduced yields of microbial genomic DNA. For convenient high-throughput 96-well homogenization, we offer the TissueLyser II (cat. no. 85300) and Plate Adapter Set (cat. no. 11990). In conjunction with PowerBead Pro Plates, the TissueLyser II provides high-throughput processing for simultaneous, rapid, and effective disruption of up to 2 x 96 samples in only a few minutes.

For disruption using 2 ml PowerBead Pro Tubes, the TissueLyser II provides simultaneous disruption of up to 48 samples in combination with the TissueLyser Adapter Set 2 x 24 (cat. no. 69982) or up to 96 samples in combination with the 2 ml Tube Holder Set (cat. no. 11993) and Plate Adapter Set. Alternately, the PowerLyzer® 24 Homogenizer (cat. no. 13155) allows the simultaneous disruption of up to 24 PowerBead Pro Tubes.

Homogenization using Vortex-Genie® 2 with Vortex Adapter for 1.5–2 ml tubes (cat. no. 13000-V1-24) can also be used for disruption in conjunction with PowerBead Pro Tubes. The order of placement of components and reagents for the platform portion of the protocol will be described in the downloaded software specific to the KingFisher platform being used.

Other open platform robots may be used with this kit. However, you may need to contact the manufacturer of your robot for help in adapting this protocol to their system.