GSM5024880: U251-overexpression MDM2 rep1; Homo sapiens; RNA-Seq
A RNeasy Mini Kit (Qiagen 74104) was used to extract total RNA. The RNA products isolated using this kit were loaded on a 1% agarose gel and separated by electrophoresis. Then, bands for 28S RNA, 18S RNA, and 5S RNA were inspected to ensure that RNA was extracted correctly. Next, the RNA quality and concentration were assayed using a Nanodrop 2000 spectrophotometer (ThermoFisher Scientific). Strand-specific libraries were generated by using the KAPA Stranded mRNA-Seq Kit with KAPA mRNA Capture Beads (KAPA Biosystem Wilmington, MA). Sequencing libraries were quantified using the Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) and validated using High-Performance Capillary Electrophoresis (HPCE) and the Agilent Tapestation 4200 (Agilent Technologies, CA, USA).he sequencing libraries were multiplexed and clustered on one lane of a Flow Cell. After clustering, the Flow Cell was loaded on the Illumina HiSeq instrument according to manufacturer's instructions.  RNA-seq was performed by GENEWIZ (Plainfield, NJ, USA)