GSM5827030: MeDIP_Agomirnc_IP; Homo sapiens; MeDIP-Seq
Total RNA was extracted from Tfh cells using TRIzol reagent (MRC) for RNA-sequencing, including total RNA sample detection, mRNA enrichment, synthesis of double-stranded cDNAs, End Repair/dA-Tailing Module, fragment selection, and PCR amplification, library detection, and Illumina sequencingselection, and PCR amplification, library detection, and Illumina sequencing. DNA was isolated from induced Tfh cells. 1 microgram of original genomic DNA was fragmented to a mean size of 300 bp. The methylated DNA fractions were immunoprecipitated using the Magnetic Methylated DNA Immunoprecipitation Kit (Diagenode) according to the instructions. The hydroxymethylated DNA fractions were immunoprecipitated using the hMeDIP kit (Active Motif) according to the instructions. After that, the immunoprecipitated products were then treated with protease K and purified with ZYMO DNA Clean & Concentrator-5 (ZYMO). Finally, the MeDIP seq and hMeDIP-seq libraries were constructed by PCR amplification with the enriched DNA as templates. RNA libraries were prepared for sequencing using standard Illumina protocols MeDIP-seq/hMeDIP libraries were sequenced using Illumina platform with PE150 read length.