16S amplicon-seq of Macrostomum lignano: food
The 16S rRNA gene primer pair 27F/338R was used to test the quality of DNA samples in a 20 l duplex PCR reaction with Phusion High Fidelity DNA Polymerase (Thermo Fisher Scientific, Waltham, USA). The proceed of PCR program as follows: a 3 min pre-denaturation step at 98 C, followed by 35 cycles (98 C for 10 s, 58 C for 30 s and 72 C for 1 min), and thereafter a final 5 min elongation step at 72 C. The PCR products were checked by 2% agarose gel, the DNA samples having clear and right signals were chosen to sequence the bacterial 16S V2 region through the MiSeq platform (Illumina).