GSM2587521: Namalwa RNA-seq rep 1; Homo sapiens; Human gammaherpesvirus 4; RNA-Seq
We isolated RNA from 5 × 10^6 log phase cells homogenized with a QIAshredder spin column (Qiagen). Total RNA was purified by silica-based membrane affinity as packaged in the RNeasy Mini Kit (Qiagen). Preparations included the optional DNase treatment step. Single primer isothermal linear amplification to cDNA was achieved using the Ovation RNA-Seq System V2 (NuGEN) and 20 ng of RNA. 3 μg of cDNA was then sheared in a 40 μL volume using a Covaris S2 Focused-ultrasonicator. We prepared deep sequencing libraries by adaptor-mediated amplification using Ovation Ultralow Library System V2 (NuGEN).