Figure 1. NucleoMag DNA Bacteria procedure. For optimal DNA yields, complete disruption of sample material is necessary and can be performed using MN bead tubes and/or MN 96 bead plates. After sample disruption, Binding Buffer IMB and NucleoMag B-Beads are added to the transferred lysate. Following magnetic separation, NucleoMag B-Beads are washed to remove contaminants and salts using Wash Buffer IMW and 80% ethanol, respectively. After air-drying the NucleoMag B-Beads for 10 min at room temperature, the DNA is finally eluted with Elution Buffer IME. The NucleoMag DNA Bacteria kit can be used either for manual processing or it can be automated on standard liquid handling instruments and magnetic separators.

Figure 2. More efficient extraction of microbial DNA than competitor kits. DNA was isolated from Gram-positive and Gram-negative bacteria as well as yeast using the NucleoMag DNA Bacteria kit (MN, blue bars) in comparison with Competitor Q (gray bars, Panel A) or Competitor T (gray bars, Panel B) according to the manufacturers' recommendations. The isolated DNA was then analyzed by qPCR analysis of 16s and 18s rRNA sequences using the Maxima SYBR Green kit by Thermo Fisher Scientific on an Applied Biosystems 7500 Real-Time PCR System. Resulting Ct values were consistently lower for DNA isolated using NucleoMag DNA Bacteria than Ct values obtained with DNA isolated using the competitor kits, suggesting that NucleoMag DNA Bacteria provided more efficient extraction of genomic DNA from the various samples.

Figure 3. Efficient DNA extraction using either bead-plate or single-tube homogenization. DNA was isolated from various Gram-positive and Gram-negative bacteria using the NucleoMag DNA Bacteria kit in combination with different bead formats for sample homogenization and resulting DNA yields were measured by UV spectrometry. The samples were homogenized by using either racks of prefilled tube strips (MN 96 bead plates) or single bead tubes (MN bead tubes). The resulting yields for samples homogenized with MN 96 bead plates (dark blue bars) were comparable to the yields obtained via homogenization in the MN bead tubes (light blue bars).

Figure 4. Reliable DNA integrity with bead-plate or bead-tube homogenization. DNA was isolated from Gram-positive and Gram-negative bacteria using the NucleoMag DNA Bacteria kit, and samples were homogenized in parallel by using either a rack of prefilled tube strips (BP = MN 96 bead plate) or single bead tubes (BT = MN bead tube). Both homogenization approaches enabled reliable DNA extraction when used in tandem with the NucleoMag DNA Bacteria kit.