Microbiota analysis of the effects of fermented blueberry pomace on laying hens

The microbial DNA from the individual cecal contents was extracted using the Fast DNA SPIN extraction kit (MP Biomedicals, Santa Ana, CA, USA) according to the manufacturer’s instructions. The V3-V4 hypervariable regions of the microbial 16S rRNA gene were amplified using PCR with the forward primer F (5′-ACTCCTACGGGAGGCAGCA-3′) and the reverse primer R (5′-GGACTACHVGGGTWTCTAAT-3′). Illumina MiSeq sequencing and bioinformatics analysis were performed by the Personal Biotechnology Co., Ltd. (Shanghai, China) on Illumina NovaSeqPE 250 platform (Illumina, San Diego, USA). In theory, each group includes eight sets of sequence data. In this dataset, the FBPM group only had seven sets of sequence data due to the death of laying hens.