Magna pure bacterial lysis buffer

DNA isolation from saliva samples was performed according to the manufacturer's guidelines (Roche, Mannheim, Germany). In brief, 200 µl of sample material was thoroughly vortexed, supplemented with 180 µl MagNA Pure Bacterial Lysis buffer (Roche) and 20 µl Proteinase K 20 mg/ml (Roche), and incubated for 1 hour at 37°C. Purification was performed in a Roche MagNA Pure 96 automated DNA extractor, using MagNA Pure 96 DNA and viral NA Small Volume kit (Roche) by specifications according to protocol Pathogen_Universal_200. The final elution volume was 50 µl. HOMIM analyses were performed as described (6 (link)). In brief, HOMIM is a molecular technique using 2 separate polymerase chain reaction (PCR) amplifications and subsequent DNA–DNA hybridization for identification of around 300 bacterial species, as single-stranded fluorescent labeled PCR products are captured by highly specific bacterial 16S rRNA probes (18–20 bases) printed on a customized aldehyde-coated glass slide. In conjunction with studies conducted on saliva samples from DANHES, all probes were thoroughly validated. Probes with cross-reactions and/or missing reactions were excluded from statistical analysis. Further analysis and generation of microbial profiles from scanned microarrays were performed with the HOMIM online tool for analysis (http://bioinformatics.forsyth.org/homim/).