GSM6222097: subject WF-035 followup; Homo sapiens; RNA-Seq
Total RNA was extracted using the miRNAeasy mini kit (Qiagen Sciences, Inc., Germantown, MD). The cDNA library preparation for total RNA from sputum cells was performed with the Illumina TruSeq Stranded Total RNA (Illumina, Inc., San Diego, CA). Libraries were quality checked by quantification with PicoGreen, size analysis on a Bioanalyzer or TapeStation 2200 (Agilent, Santa Clara, CA) and qPCR quantitation against a standard curve. 75 bp paired end reads were generated on a HiSeq 2500 flow cell. Libraries are loaded at an empirically determined concentration in order to generate the optimal number of clusters per lane of the flow cell. Samples were sequenced with an average depth of 20 million reads using Illumina HiSeq 2500 GPL27467.