· Thermal mixer containing a 1.5 ml tube block (optional: 2 ml tube block for elution)
· Recommended: vertical rotating mixer (e.g., Thermo Scientific‱ HulaMixer‱ Sample Mixer).
· Cold PBS, 300 µl per sample (Low Input: 150 µl per sample). Alternatively, TE or Tris buffer can be used.
· Isopropanol, 550 µl per sample (Low Input: 275 µl/sample).
· 1.5 ml DNase-free, low DNA binding microfuge tubes (e.g., Eppendorf‱ DNA LoBind‱, #0030108051) are recommended for elution and storage (1 per sample); it is especially important to use low DNA binding tubes if working with UHMW DNA, which tends to bind to plastic surfaces.
· For Gram-negative bacteria: Lysozyme (25 mg/ml, 10 µl per sample)
· For Gram-positive bacteria: STET Buffer (Current Protocols in Molecular Biology) containing Lysozyme (10 mg/ml) can be an effective lysis agent (150 µl or 300 µl per sample).
· Additional lysis agents may be required (e.g., lysostaphin).