Transcriptome Analysis of Lin- and Lin+ Fractions

RNA was isolated from Lin− and Lin+ fractions (25-50 x 10³ cells, respectively) after FACS. RNA extraction was performed using a MicroRNA Isolation kit (Qiagen Inc., Cat#74004, Germantown, MD). 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, Cat#634946, San Jose, CA). The Ion Plus Fragment Library kit (Thermo Fisher, Cat#4471252) was used for fragmented RNA, as previously reported (30 (link)–32 (link)). The Ion Proton S5/XL platform (Thermo Fisher) was used for sequencing at the Analytical and Translational Genomics Shared Resource Core at the University of New Mexico Comprehensive Cancer Center (UNM-CCC).