MethylTarget™ assays were performed by Genesky BioTech (Shanghai, China) [12 (link), 23 (link)]. Briefly, we used an EZ DNA Methylation™-GOLD Kit (ZYMO RESEARCH) to transform all unmethylated cytosines to uracils. The samples with a bisulfite conversion rate < 98% were first filtered out. After the target CpG regions were amplified, separated, and purified by a gel extraction kit (Vazyme Biotech), a CpG island methylation assay was performed with an Illumina Hiseq/Miseq 2000 according to the manufacturer's protocol.
Hiseq miseq 2000
Manufactured by Illumina
Sourced in United States
The Illumina HiSeq/MiSeq 2000 is a high-throughput DNA sequencing platform. It utilizes sequencing-by-synthesis technology to generate large volumes of sequence data. The system is capable of producing millions of DNA sequence reads in a single run.
Automatically generated - may contain errors
2 protocols using hiseq miseq 2000
1
Methylation Profiling by Illumina Sequencing
2
Gut Microbiome Profiling by NGS
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The stored gut parts were crushed and powdered in liquid nitrogen. Total genomic DNA was extracted using the Wizard® Genomic DNA Puri cation kit (www.promega.com) and shipped to Beijing Genomics Institute, China for next generation sequencing. Based on the PCR results, a high-quality original library was prepared by Illumina kit by removing short fragments. Illumina (USA) HiSeq/MiSeq 2000 encoded 16S rRNA gene amplicons were used to observe the gut microbiome diversity (19) . The quality of the raw data sequences was checked using FastQC v0.11.9. Filtration of the raw data was done to obtain clean reads, and tags were clustered at 97% sequence identity to an operational taxonomic unit (OTU), which assigned its taxonomy.
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