16S_of_human_gut_community
We amplified the V3 and V4 region of the 16S rRNA gene with three slightly modified universal primers60 with an increment of a 1-nt frameshift in each primer to increase MiSeq sequencing output performance between target region and Illumina adapter. The target region was amplified using 4 primer pairs in a limited 17-cycle PCR and performing 4 different PCR reactions per each primer pair combination separately.