GSM6543111: 4:1_1; Homo sapiens; RNA-Seq
Total RNA was extracted from spheroids by using TRIzol reagent (Invitrogen). 0.9 ug of RNA was used for constructing sequencing libraries. Following the manufacturer's instructions, the library was constructed by using a QuantSeq 3′ mRNA-Seq Library Prep Kit (Lexogen, Austria).  Each total RNA were prepared and an oligo-dT primer containing an Illumina-compatible sequence at its 5' end was hybridized to the RNA and reverse transcription was performed. After degradation of the RNA template, second strand synthesis was initiated by a random primer containing an Illuminacompatible linker sequence at its 5' end. The double-stranded library was purified by using magnetic beads to remove all reaction components. The library was amplified to add the complete adapter sequences required for cluster generation. The finished library is purified from PCR components.