16S rRNA gene sequencing of microbiota in human gastrointestinal biopsies - standards
The human microbiota is currently developing field of research. For its study, the hypervariable regions (V) of 16S ribosomal RNA gene (16S rDNA) sequencing in combination with Illumina patforms are widely used. However, the primer design targeting the V regions without the off-target amplification of abundantly presents human DNA in tissue samples is challenge. In our metagenome analysis of upper gastrointestinal tract biopsies, we observed non-specific amplification of human DNA in case of using 515F-806R primers targeting V4 region. Therefore, we designed specific set of V1-V2 primers that in combination with MiniSeq platform guarantee optimal microbiota resolution for microbial low-abundant samples without human off-target amplification.