Fig. 3

Cas-16S-seq efficiently removed the rice mitochondrial 16S rRNA gene fractions in mock communities. Three gRNAs (mt-gRNA1048, mt-gRNA1171, and mt-gRNA1196) were used to evaluate the performance of Cas-16S-seq. Mock samples containing 0%, 0.1%, 0.2%, 20%, 40%, 50%, and 100% bacterial 16S rRNA gene fragments were treated with Cas9/gRNA and then amplified by PCR. The depletion efficiency of host 16S rRNA genes was estimated by gel electrophoresis since the mitochondrial PCR product is approximately 84 bp larger than that of bacteria. The PNA PCR clamping method was also performed using the same mock samples. Arrows indicate the treatments by which the mitochondrial 16S rRNA gene fraction was efficiently eliminated. M, DNA marker; bp, base pair