Profiling RNA Expression in Hematopoietic Cells

RNA was isolated from 25–50 × 10³ cells from the Lin− and Lin+ fractions after FACS using a MicroRNA kit (Qiagen, Cat# 74004, Germantown, MD, USA). RNAs from matching Lin− and Lin+ patient samples were also compared with RNA from PBMC samples of normal healthy donors (negative controls). Synthesis of cDNA and library preparation were performed using the SMARTer Universal Low Input RNA kit for sequencing (Clontech, Cat# 634946, San Jose, CA, USA), and the Ion Plus Fragment Library kit (Thermo Fisher, Waltham, MA, USA, Cat# 4471252), as previously described [25 (link),26 (link),27 (link)]. Sequencing was performed using the Ion Proton S5/XL system (Thermo Fisher, Waltham, MA, USA) in the Analytical and Translational Genomics Shared Resource at the University of New Mexico Comprehensive Cancer Center.