The effect of Zhejiang psyllium extract on gene expression of gut microbiota in hyperuricemia rats
Genomic DNA was extracted from fecal samples using the CTAB method and the extracted genomic DNA was detected using 1% agarose gel electrophoresis. The instrument Covaris M220 was used to fragment the DNA to a size of 350 bp. A PE library was constructed by attaching specific adapters to both ends of the DNA fragments, and magnetic beads were used to screen and remove adapter-self-ligated fragments. PCR amplification was performed to enrich the library template, followed by sodium hydroxide denaturation to produce single-stranded DNA fragments. Bridge PCR was conducted to amplify the DNA molecules, ensuring high efficiency and accuracy during sequencing. Illumina sequencing was performed as required, and the sequence of the template DNA fragments was determined by collecting the fluorescence signals in each cycle. For data analysis, the raw sequences were first optimized through splitting, quality trimming, and contamination removal. The optimized sequences were then used for assembly and gene prediction. The obtained genes were annotated and classified in terms of species and function to obtain valid data for subsequent analysis.