Fecal slurry ISO equol producer WP, replicate 1
This sample was prepared with feces from a menopausal woman previously classified as equol producer. Ten-fold dilution of this feces was used to inoculate with 10% (w/v) a fermentation flask containing sterile modified medium for colon bacteria (Van der Meulen et al., 2006) with decreased glucose content (2g L-1) and with isoflavone supplementation (160 mg L-1 isoflavones Fisiogen, Zambon) (MCBISO). After incubation with magnetic stirring (250 rpm) for 24 h in anaerobiosis at 37 °C the fecal slurry was centrifuged at 13,200 rpm 10 min and pellets were collected. Extraction of total bacterial DNA was performed based on a phenol-chloroform protocol described by Zoetendal et al. (2006) in duplicate. This experiment includes sequencing of one replicate of DNA extraction. The 16S rRNA gene was amplified with primers 515F and 909R (targeting bacterial 16S Region V4-V5) (Li et al., 2014) including Illumina adapters and paired-end sequenced in an Illumina Miseq System (StarSeq, Mainz, Germany)