human oral metagenome
Root Canal Sample collected from the root canal space of the diseased tooth.  After rubber dam isolation, the tooth surface was disinfected with 30% hydrogen peroxide, followed by 5% tincture of iodine, 5.25% sodium hypochlorite and finally 5% sodium thiosulfate to inactivate the halides. The pulp chamber was accessed with #4 round carbide bur followed by an endo Z bur. Canals were identified under a surgical operating microscope. Three Fine paper points were placed in each canal to collect fluids from the canal space.  If purulence or serous fluid was present in the canal, this was directly sampled with three fine size paper points followed by a sterile file to the working length, as indicated by an apex locator and then aseptically separated into the sampling vial. In multicanaled teeth, one paper point sample was obtained from each canal unless the canals were very calcified, in which case sampling of the canal root with the largest periapical lesion and the largest canal was sampled. If the canal was dry, sterile saline was added, the biofilm disrupted with a file, and the file together with three paper points were similarly sampled. The file and paper points were placed in sterile, DNA- and RNA-free vials containing 1.5mL of filter sterilized 10mM Tris-HCl, (pH 8.0).