Sample preparation

After removal of supragingival plaque above the gingival line, isolate the sampling sites with sterile cotton roll, and then air-dry.

Remove the subgingival plaque samples from the deepest pockets from the same patients with individual sterile Gracey curettes.

Immerse the plaque sample in 1.0 ml PBS (-) (GibcoBRL, MD, USA), and vortex for 30 sec, followed by centrifugation for 20 min at 15,000 g. Remove the supernatant.

Extract the bacterial DNA from cultivated strains and clinical plaque samples by using InstaGene® Matrix (Bio-Rad Lab., CA, USA), according to manufacturer's instructions.

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