Chunk #36 — Experimental Procedures — RNA Isolation and qRT-PCR

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In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling.

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Total RNA was isolated from differentiated cells using Qiazol (Qiagen) according to the manufacturer's instructions. Reverse transcription was performed with the SuperScript III first-strand synthesis system (Life Technologies). qPCR was performed using Power SYBR Green PCR Master Mix (Applied Biosystems) and a StepOnePlus system (Applied Biosystems). The amount of target RNA was normalized relative to the amount of 18S rRNA mRNA. Primer sequences are shown in Table S1.