title: GSM8301423: rhLUM-C; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from MHCC97L xenograft using RNAiso Plus (Takara Bio Inc., Kusatsu, Shiga, Japan) according to manufacturer's protocol. Quality and quantity of total RNA samples were assessed using the RNA 6000 Nano total RNA assay (Agilent) by Agilent 2100 bioanalyzer and Qubit 2.0 Fluorometer, respectively. Qualified RNA (1 μg) samples were subjected to library preparation using KAPA mRNA HyperPrep Kit (KR1352). Poly-A containing mRNA was collected by using poly-T oligo-attached magnetic beads. The purified mRNA was fragmented to 200 – 300bp by incubating at 94oC for 6 min in the presence of magnesium ions. The fragmented mRNA was then applied as template to synthesize the first-strand cDNA by using random hexamer-primer and reverse transcriptase. In the second strand cDNA synthesis, the mRNA template was removed and a replacement strand was generated to form the blunt-end double-stranded (ds) cDNA. The ds cDNA underwent, 3' adenylation and indexed adaptor ligation. The adaptor-ligated libraries were enriched by 10 cycles of polymerase chain reaction (PCR). The libraries were denatured and diluted to optimal concentration. Illumina NovaSeq 6000 was used for Pair-End 151bp sequencing.