This sample kit is optimized to make evaulation of the PowerMag® Soil DNA Isolation Kit easy. In the sample kit only, the homogenization and inhibitor removal steps are performed manually in 2 ml tubes instead of in 96 well plates. This allows the kit to be tested without the use of a 96 well plate shaker. Following the inhibitor removal step, samples are transferred to the deck of a KingFisher® Flex or KingFisher® Duo for automated DNA purification.
The PowerMag® Soil DNA Isolation Kit is designed for automated isolation of microbial DNA from all soil types and other environmental samples, as well as stool, compost, sediment and manure. The protocol is designed for isolation of DNA from up to 0.25 g of sample and includes patented Inhibitor Removal Technology® to remove humic substances and other PCR inhibitors. Novel magnetic particle technology enables purification of DNA without the typical surface binding to the beads, eliminating the adsorption of organic inhibitors that is typical of other magnetic bead technologies, and facilitating isolation of pure DNA. In addition, neither chaotropic salts nor ethanol are used in the binding and washing steps, removing a second source of contamination that can inhibit enzymatic reactions. The PowerMag® Soil DNA Isolation Kit has been optimized for use with the Thermo Scientific KingFisher® Flex and KingFisher® Duo automated processing systems, for rapid, high-throughput isolation of inhibitor-free DNA. The high-quality DNA isolated with the PowerMag® Soil DNA Isolation Kit is ready to use in PCR, qPCR and next generation sequencing.
|Figure 1. High quality, pure DNA isolated from a panel of six soil samples (0.25 g) using the PowerMag® Soil DNA Isolation Kit on the KingFisher® Duo. High quality, high molecular weight DNA was observed in each soil sample with yields varying based on microbial load. No differences in yield or quality were observed between the replicate samples. qPCR was performed on 1 µl of DNA from three soil types known to be high in PCR inhibitors, using primers for Bacillus. Undiluted DNA and samples diluted 1:10 fell within the standard curve (grey lines), and the difference between the diluted and undiluted samples was approximately 3 cycles, indicating the DNA was free of inhibitors.|
|Figure 2. Inhibitor Removal Technology® (IRT) is essential for removal of PCR inhibitors. DNA was isolated from organic garden soil samples using the PowerMag® Soil DNA Isolation Kit with and without the IRT step and purified using the KingFisher® Duo. qPCR was performed as described in Figure1. Samples processed without IRT failed to amplify, while samples with IRT were successfully amplified and fell within the standard curve (blue lines).|
|Starting Amount||0.25 g|