16S mcirobiome analysis of ileum from Ceftriaxone treated mice, D22
Procedures were performed as previously described in detail. Genomic DNA was extracted using DNeasy Blood and Tissue Kit (Qiagen, 69504) and homogenized in bead beating tubes (Revvity, 19-624). Raw cecal slurries were processed in duplicate. The V4 region of the 16S rRNA gene was amplified according to protocol, and sequencing was performed on the Illumina MiSeq platform (San Diego, CA) as previously described. AccuPrime High-Fidelity Taq was used, with PCR cycling conditions as follows: 95C for 2 min, then 20 cycles of touchdown PCR followed by 20 cycles of standard PCR, and finally at 72C for 10 min. Sterile water, DNA isolation controls, and empty wells were used as negative controls. Synthetic standard bacterial communities (Zymo Research, Irvine, CA; catalog no. D6306) were used as positive controls.
SUB16152516
SLjT2_Ileum_1