GSM8817999: D11 UHRF1fl/fl iTreg 2; Mus musculus; OTHER
Genomic DNA was extracted from flow cytometry-sorted cells using the AllPrep DNA/RNA Micro Kit. Genomic DNA was quantified with a Qubit 3.0 instrument.  gDNA was fragmented with MspI (New England Biolabs) and then size-selected for 100- to 250-bp fragments using solid-phase reversible immobilization beads (MagBio Genomics). CG-enriched gDNA was then enzymatically converted using the NEBNext Enzymatic Methyl-seq Kit (New England BioLabs) per manufecture's instruction. Random priming, adaptor ligation, PCR product clean-up, and final library amplification were performed using the Pico Methyl-Seq Library Prep Kit (Zymo Research). Libraries were run on a High Sensitivity chip using an Agilent TapeStation 4200 to assess size distribution and overall quality of the amplified library. Fluorometric quantitation and TapeStation size distribution estimates permitted equimolar pooling, and 4-6 pooled libraries per run were sequenced on an Illumina NextSeq 2000 instrument using the 100 cycles NextSeq 2000 P2 reagent kit.