title: 16S rRNA of mus musculus:Pla2g10+/+ mice chyme
design: Genomic DNA (gDNA) was extracted from the colonic chyme using a soil genomic DNA extraction kit (#DP336, Tiangen biotech, Beijing, China). The gDNA was used as a template to amplify the highly-variable V3V4 region of the 16S RNA gene by PCR, using the forward primer 341F (5-ACTCCTACGGGAGGCAGCAG-3) and the reverse primer 806R (5-GGACTACHVGGGTWTCT AAT-3). All PCRs were performed in 15L of Phusion High-Fidelity PCR Master Mix (New England Biolabs, Manchester, UK), with 0.2 M of both the forward and reverse primers, and approximately 10 ng of template DNA. Thermal cycling consisted of an initial denaturation at 98C for 1 minute, followed by 30 cycles of denaturation at 98C for 10s, annealing at 50C for 30s, and elongation at 72C for 30s, and finally an extension at 72C for 5 min.
library_name: FKDO230571947-1A