Mouse primary neuron RNA extraction and real-time PCR

Total RNA was extracted from cell lysates using TRIzol Reagent (Invitrogen, Carlsbad, CA), and 1 μg was retrotranscribed using the ThermoScript RT-PCR System (Invitrogen, Carlsbad, CA). Real-time PCRs were performed in triplicate on an Applied Biosystems 7900HT Fast Real-Time PCR system using 10 ng of cDNA, 5 μl of SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA), and 150 nM specific primers (sequences available on request) in a final volume of 10 μl.

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