GSM7034841: SS028; Homo sapiens; RNA-Seq
Blood samples were collected  and irradiated using a Clinac at dose rates between 1Gy/min and 10^5 Gy/sec (total dose 3Gy). After overnight freezing, the samples were thawed at room temperature and left for > 2 h before processing for total RNA isolation using the PAXgene® Blood RNA kit (Qiagen, Valencia, CA) according to the manufacturer's protocol. Isolated RNA was quantified using the Nanodrop One spectrophotometer (Thermofisher) and A260/ 280 ratios recorded. RNA sequencing analysis for gene expression was performed according to standard RNA-seq pipeline at the Columbia University Medical Center Genome Center. We used the RIBOZERO workflow in which ribosomal depletion was performed to remove rRNAs from globin cleared total RNA. RINs of globin cleared RNA (catalog number AM1981 from Thermofisher) were in the acceptable range for 40-million reads RNAseq (75% of samples had RIN values >7.5). After rRNA depletion, library construction was performed using Illumina TruSeq Stranded Total RNA kit.