GSM8758131: 16s microbiome data for patient 12 as EBS sample at week 12, rep 1; Homo sapiens; OTHER
Genomic DNA was extracted using a Maxwell 16 LEV Blood DNA Kit (Promega, Madison, WI) implemented on a Maxwell 16 Instrument, following the manufacturer's instructions with minor modifications Genomic DNA was prepared for sequencing on a PacBio Revio sequencer using a two-stage PCR protocol, similar to that described for Illumina amplicon library preparation.15 In the first stage of PCR, DNA was PCR amplified with primers 27F and 1492R (TTTCTGTTGGTGCTGATATTGCAGRGTTYGATYMTGGCTCAG and ACTTGCCTGTCGCTCTATCTTC RGYTACCTTGTTACGACTT, respectively) using domain-level bacterial primers as described by PacBio. The underlined regions, however, represent Oxford Nanopore Universal sequences for tailing PCR primers' linker sequences. In the second stage of PCR, amplicons from the first stage were used as a template and amplified with primers that contained PacBio Kinnex adapter sequences, combinatorial dual indices, and the ONT linker sequences at the 3' ends of the primers . Library preparation was performed using a two-stage PCR amplification protocol but with primers targeting the full 16S rRNA gene