Intestinal microbiota of 18-day-old broiler raw sequence reads
DNA was extracted from the crop, ileal and cecal digesta (~250 mg) and the V4 region of bacterial 16S rRNA gene was amplified. PCR amplification was performed using universal primers. The amplification program consisted of one cycle of 94 °C for 3 min; 40 cycles of 94 °C for 45 sec, 55 °C for 1 min, and 72 °C for 1.5 min; and finally one cycle of 72 °C for 10 min. Amplicons were pooled together, and DNA library was constructed using NEBNext Ultra DNA Library Prep Kit for Illumina (New England BioLabs, MA, USA).