NovaSeq of mouse:6_2_6Z42
Bacterial samples obtained from either in vitro cultures or organ homogenates were plated on THB agar plates containing Kan (150 g/mL) and incubated overnight at 37 . Bacteria were harvested from the plates using PBS and subsequently stored at 80 . Genomic templates were prepared by diluting bacterial samples in water (1:50) and boiling at 95 for 15min. 1 l of the sample was used as the template for PCR amplification (with primers P5/P7), and cycling conditions for PCR were the following: initial denaturation 95 300 s, 27 cycles of 95 15 s, 55 15 s, and 72 10 s, and final extension of 72 5 min. Reactions were carried out with a total volume of 50 l using 2 Rapid Taq Master Mix (Vazyme, China). The presence of amplicons was pooled, verified and purified by agarose gel extraction (Omega, USA). Purified amplicons were quantified by Qubit dsDNA HS Assay Kit (Thermo Fisher, USA) and sequenced with NovaSeq (Illumina).