title: The V3 and V4 region genes of 16SrRNA of cecal microorganisms in male broilers
design: Genomic DNA was extracted with the cetyltrimethylammonium ammonium bromide (CTAB) method. The purity and concentration of total DNA were detected by agarose gel electrophoresis. Amplicons from the V3 and V4 regions of the 16S rRNA gene were generated from the extracted DNA using the primers 341F (5-CCTAYGGGRBGCASCAG-3) and 806R (5-GGACTACNNGGGTATCTAAT-3). The sequencing libraries were constructed by NEBNext Ultra IIDNA Library Prep Kit and quantified by Qubit and Q-PCR. After being confirmed qualified, NovaSeq6000 was used for machine sequencing. Demultiplexed raw sequence data were quality-filtered and denoised using DADA2 method of QIIME2 to generate amplicon sequence variants (ASVs).