16S rRNA sequencing of human: breast milk day 15
DNA was extracted using Milk DNA Preservation and Isolation Kit (Cat. 44800, Norgen Biotek). The V4-V5 region of the 16S rRNA gene was amplified using primers 515F (5-GTGYCAGCMGCCGCGGTAA-3) and 926R (5-CCGYCAATTYMTTTRAGTTT-3). Amplicons were normalized with the Just-a-Plate 96-well normalization kit (Charm Biotech).