RREM-seq (reduced representation enzymatic methylation sequencing) of Homo sapiens: flow-sorted Tconv cells from bronchoalveolar lavage fluid
Immediately after sorting, cells were pelleted by centrifugation and lysed in 350 μl RLT Plus lysis buffer (Qiagen) supplemented with 2-mercaptoethanol. Lysates were stored at −80 °C until DNA isolation using the AllPrep DNA/RNA Micro kit according to the manufacturer's protocol (Qiagen). DNA quantity was assessed using Qubit dsDNA HS Assay Kit, and RREM-seq libraries were prepared from 1-2ng  of DNA using our custome RREM-seq protocols. Libraries were pooled using dual indexing and sequenced on a NextSeq 2000 instrument (Illumina), single-end.