TruSeq Stranded mRNA

Protocol tips

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	The protocol has been tested using 0.1–1 µg of high-quality universal human reference total RNA as input.

Protocol tips
The protocol has been tested using 0.1–1 µg of high-quality universal human reference total RNA as input.

Publication protocol

PANC-1 (0.35 × 106 cells/well) cell line was seeded overnight in 60 mm plates. Next day, cells were treated with 400 nM CM03 for 6 and 24 h. For gemcitabine treatment, cells were treated for 6 and 24 h with DMSO or 30 nM, 100 nM, 300 nM, 1 μM, 3 μM, and 10 μM gemcitabine-HCL for PANC-1 cells and 30 nM, 300 nM, and 3 μM gemcitabine-HCl for MIA PaCa-2 cells. Total RNA was extracted using the Qiagen RNeasy Plus Mini Kit (ThermoFisher, catalogue no. 74134) as per the manufacturer’s instructions. RNA quality (RIN > 9.0) was checked with an Aligent 2100 bioanalyzer RNA 6000 Nano Chip, and RNA concentration was quantified using a Qubit fluorometer (ThermoFisher) and Qubit RNA HS Assay Kit (Qiagen, catalogue no. Q32852). RNA-seq libraries were then generated using the Illumina Truseq RNA HT (stranded mRNA) kit (Illumina, catalogue no. RS-122–2103) as per the manufacturer’s instructions and sequenced using an Illumina HiSeq4000.

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Manufacturer protocol

Download the product protocol from Illumina for TruSeq Stranded mRNA below.

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