Six-year-old lactating donkey 16SrDNA: rectal feces
Firstly, sample DNA was extracted, specific primers with barcode in the assay region were synthesized, and the total DNA of the extracted samples was used as template to select specific primers for bacterial 16S rRNA and fungal ITS rRNA high variation regions for PCR amplification using an ABI GeneAmp 9700 PCR instrument. The PCR products of the same samples were mixed and detected by 2% Agarose gel electrophoresis was used to detect the PCR products, and the PCR products were recovered by cutting the gel using the AxyPrepDNA gel recovery kit (AXYGEN) and eluted with Tris-HCl; 2% agarose electrophoresis was used to detect the PCR products. The PCR products were detected and quantified by QuantiFluor-ST Blue Fluorescence Quantification System (Promega), and then the samples were sequenced using Illumina Miseq sequencing platform.