GSM7232610: BT474 Vehicle 007; Homo sapiens; RNA-Seq
Tumors were dissected and a portion was placed into RNALater and frozen at -70. Total RNA was extracted using Qiagen RNeasy Plus Universal mini kit following manufacturer's instructions (Qiagen, Hilden, Germany). The RNA samples received were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked using TapeStation (Agilent Technologies, Palo Alto, CA, USA). Prep Kit for Illumina using manufacturer's instructions (NEB, Ipswich, MA, USA). The RNA sequencing library was prepared using the NEBNext Ultra RNA Library The sequencing library was validated on Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA)  and quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). The sequencing library was clustered on one lane of a flowcell. The flowcell was loaded on the Illumina HiSeq instrument (4000 or equivalent) according to manufacturer's instructions. The sample was sequenced using a 2x150bp Paired End (PE) configuration. Image analysis and base calling were conducted by the HiSeq Control Software (HCS).