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 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.