RNA-Seq of Lin+ and Lin- Fractions

RNA was isolated from Lin− and Lin+ fractions (25–50 × 10³ cells, respectively) after FACS. RNA extraction was performed using a miRNA Isolation kit (Qiagen Inc., catalog no. 74004). RNA from matching Lin− and Lin+ fractions were compared with RNA from PBMCs of healthy donors (negative controls). RNA analysis, cDNA amplification, and library preparation were performed using the human microarray platform (SMARTer Universal Low Input RNA kit for sequencing (Clontech, catalog no. 634946). The Ion Plus Fragment Library kit (Thermo Fisher Scientific, catalog no. 4471252) was used for fragmented RNA, as reported previously (30–32 (link)). The Ion Proton S5/XL platform (Thermo Fisher Scientific) was used for sequencing at the Analytical and Translational Genomics Shared Resource Core at the University of New Mexico Comprehensive Cancer Center (UNM-CCC).