Bacterial 16S Ribosomal RNA Profiling

Partial 16S rRNA gene sequences were amplified from the bacterial DNA extracted from stool samples using primer pair Probio Uni and/Probio_Rev, which targets the V3 region of the 16S rRNA gene sequence27 (link). Illumina adapter overhang nucleotide sequences were added to the 16S rRNA gene-specific sequences. The 16S rRNA gene amplicons were prepared following the 16S Metagenomic Sequencing Library Preparation Protocol. Amplifications were carried out using a Verity Thermocycler (Applied Biosystems). The integrity of the PCR amplicons was analysed by electrophoresis on a 2200 TapeStation Instrument (Agilent Technologies, USA).