High-throuhgput 16S rRNA sequencing from mice gut
The conserved regions V3 and V4 (459 bp) of the 16S rRNA gene were then amplified using forward and reverse primers: 5-TCG TCG GCA GCG TCA GAT GTG TAT AAG AGA CAG CCT ACG GGN GGC WGC AG 3 and 5- GTC TCG TGG GCT CGG AGA TGT GTA TAA GAG ACA GGA CTA CHV GGG TAT CTA ATC C -3, respectively. Amplification was carried out using the KAPA HiFi HotStart ReadyMix PCR kit (KK2602) and the following PCR cycle. Amplicons were mixed with Illumina sequencing adaptors and dual-index barcodes (Nextera XT index kit v2, FC-131-2001). Libraries were normalized and merged before the sequencing.