In this method, an extra step that consists of lysozyme-based enzymatic lysis and beating of zirconia-silica beads (BioSpec, Bartlesville, OK) was applied prior to the usage of DNeasy® Blood and Tissue kit (Qiagen Valencia, USA) as described above. Samples were transferred into clean bead beating tubes (2 ml Eppendorf tube), and 50 μl of lysozyme (20 mg/ml, Sigma-Aldrich, USA) was added to a 500 μl aliquot of cell suspension followed by incubation for 1 hr at 37 °C. Then, 600 mg of 1 mm diameter zirconia-silica beads (BioSpec, Bartlesville, USA) were added to the lysate and the cells were subjected to bead beating using a Qiagen Tissue Lyser LT at 36 Hz for 3 min. Further isolation and purification of the total genomic DNA from lysates were conducted using the DNeasy® Blood and Tissue kit according to the manufacturer’s instructions48 (link),49 (link),51 (link).
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