bacterial 16S rRNA gene V1-V9 amplicon-based microbiome profile of dental-implant-associated biofilms and sulcus fluid in a human patient
Sterile paper points were inserted into the peri-implant sulcus or pocket at six sites around each implant for 30 sec. Additionally, subgingival plaque was collected at these sites using periodontal curette and was transferred to the paper points before pooling them for each implant. RNA and DNA were simultaneously isolated from each sample using a custom protocol. Bacterial full-length 16S rDNA genes were amplified using the bacteria-specific primer pair 27F (AGRGTTYGATYMTGGCTCAG) and 1492R (RGYTACCTTGTTACGACTT). Preparation for sequencing, including ligation of barcoded SMRTbell Adapters, followed PacBio protocols.