Gut microbiota composition associated with stool consistency

Stool DNA was isolated using AllPrep DNA/RNA Mini Kit (Qiagen; cat. #80204), and subsequently we performed 16s rRNA gene sequencing using forward primer 515F (GTGCCAGCMGCCGCGGTAA) and reverse primer 806R (GGAC TACHVGGGTWTCTAAT), similar to the study by Vandeputte et al. 1 As a reference for operational taxonomic unit (OTU) picking and taxonomy annotation, we used a primer-specific version of GreenGenes 13.8, as described previously. 3 The association analysis of microbiota to BSS was corrected for sequence depth, age, gender, body mass index (BMI) and drugs that evoke an effect on the microbiome (ie, antibiotics and proton pump inhibitors).