Small RNA Sequencing of Macrophages

Small RNA libraries were prepared from size fractionation of 10ug of total macrophage RNA on a denaturing polyacrylamide gel to purify 18–35nt small RNAs, and converted into Illumina sequencing libraries with the NEBNext Small RNA Library kit (NEB). Libraries were sequenced on the Illumina NextSeq-550 in the Boston University Microarray and Sequencing Core. We applied long RNAs and small RNAs RNAseq fastq files to a transposon and small RNA genomics analysis pipeline previously developed for mosquitoes (58 (link), 106 (link)) with the mouse transposon consensus sequences loaded instead. Mouse transposon consensus sequences were downloaded from RepBase (107 (link)), and RNA expression levels were normalized internally to each library’s depth by the Reads Per Million counting method.