Chunk #43 — Methods — Illumina transcriptome library preparation and sequencing

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Characteristic analyses of a neural differentiation model from iPSC-derived neuron according to morphology, physiology, and global gene expression pattern.

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The total RNA purified from cells on neuronal differentiation day 0, day 7, and day 28 was subject to RNA-Seq analysis. RNA concentration was measured with Nanodrop One (ThermoScientific), and confirmed with Qubit 3.0 (invitrigon). The RNA integrity number was detected with BioAnalyzer 2100 (Agilent), Library construction was strictly followed Truseq RNA Access libarary Pre Kit (illumine, RS-301-2001), mRNAs were isolated from 400ng total RNA by PolyT capture beads in our lab protocol. The library length was detected by BioAnalyzer 2100 (Agilent), and the RNA concentration was confirmed by StepOnePlus™ Real-Time PCR System (Thermo fisher, 4376600). The mRNAs were sequenced with Hiseq X Ten (Illumina).