16 S rDNA of microbiome on mini-screw
The 16S rDNA hypervariable V3-V4 region was amplified by adding the primer 341F (5- CCTACGGGRSGCAGCAG -3) and 806R5'- GGACTACVVGGGTATCTAATC -3'.Unique index and adapter sequence was added to the 5' to distinguish each sample. Then PCR was performed using the KAPA HiFi HotStart kit (Kapa Biosystems, Wilmington, USA) and amplicons were purified using the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, U.S.) and the library was quantified by Qubit with the same proportion, then sequenced on the MiSeq (Illumina) PE250 platform.