GSM5170618: macrophage WT2_mock_csRNAseq; Homo sapiens; OTHER
Total RNA was extracted using Zymo Quick-RNA microprep kit. Small RNAs of ~20-60 nt were size selected from 0.4-1 µg of total RNA by denaturing gel electrophoresis. A 10% input sample was taken aside and the remainder enriched for 5'-capped RNAs with 3'-OH. Monophosphorylated RNAs were selectively degraded by Terminator 5´-Phosphate-Dependent Exonuclease (Lucigen). Subsequent 5'dephosporylation by CIP (NEB) followed by decapping with RppH (NEB) augments Cap-specific 5'adapter ligation by T4 RNA ligase 1 (NEB). The 3' adapter was ligated using truncated T4 RNA ligase 2 (NEB) without prior 3' repair to select against degraded RNA fragments.  Following cDNA synthesis, libraries were amplified for 14 cycles and sequenced SE75 on the Illumina NextSeq 500.
GEO
GSM5170618