Gansu zokor gut microbiota sequencing
Genomic DNA from gut contents was extracted (E.Z.N.A. Stool DNA Kit), checked by gel electrophoresis and NanoDrop, and used to amplify the 16S rRNA V3V4 region with primers 341F/806R. PCR (20 L: Fast Pfu buffer, dNTPs, primers, polymerase, 10 ng DNA) cycled: 95C/5 min; 30(95C/30 s, 58C/30 s, 72C/45 s); 72C/10 min. Amplicons were gel-purified (AxyPrep), quantified (Qubit 3.0), pooled equimolarly, and used to construct a sequencing library (NEBNext Ultra).