16S DNA-seq joey kangaroo stool
Briefly, the culture sample was centrifuged and freeze-dryed overnight. The next day frozen pellet was treated with a beadbeater and 750L of CTAB buffer ( 2%CTAB, 100 mM Tris 8, 20 mM EDTA, 1.4M NaCl).Phenol:Chloroform solution was used to isolate DNA used for generating amplicons that cover V3 and V4 hypervariable regions of bacteria and archaea16S rDNA. Indexed adapters were added to the ends of the 16S rDNA amplicons by limited cycle PCR.The pooled DNA libraries were loaded on an Illumina MiSeq instrument.