GSM5929313: PBMC_SIMPLE; Homo sapiens; OTHER
The formaldehyde fixed cells are collected and re-suspended in ice cold NIB buffer for nucleosome depilation; the nuclei pellets were tagmentated with identical barcoded assembled Tn5 followed 2 rounds of Ligation-based combinatorial barcoding; nuclei were lysed and gDNA purified with 1X SPRI beads; next, performs Second adaptor tagging and 5hmC transition and hmC-to-T recording; after the primer extension reaction, the dsDNA is subjected into 5mC specific labelling; the final library is amplified by Kapa HiFi uracil plus DNA polymerase and used 1.8X Ampure XP beads to purify the product. The final libraries were sequenced using a MGI-2000 platform (MGI) with the following read lengths: PE 150 + 8 + 150 (Read 1 + Index 1 + Read 2).