RNA was prepared from 106 primary mouse neuronal cells using the MagMax Total RNA isolation kit (Life Technologies). RNA (200 ng) was reverse transcribed using the High Capacity cDNA kit (Life Technologies). TaqMan gene expression assays for MT1A and TUBA8 were purchased from Life Technologies (Mm00496660_g1 and Mm00833707_mH, respectively) and qRT-PCR was performed as previously described [15 (link)]. Delta Ct method was used for normalization of expression relative to β-tubulin.
Grubman A., Lidgerwood G.E., Duncan C., Bica L., Tan J.L., Parker S.J., Caragounis A., Meyerowitz J., Volitakis I., Moujalled D., Liddell J.R., Hickey J.L., Horne M., Longmuir S., Koistinaho J., Donnelly P.S., Crouch P.J., Tammen I., White A.R, & Kanninen K.M. (2014). Deregulation of subcellular biometal homeostasis through loss of the metal transporter, Zip7, in a childhood neurodegenerative disorder. Acta Neuropathologica Communications, 2, 25.
Normalization method (Delta Ct method, relative to β-tubulin)
controls
No positive or negative controls explicitly mentioned
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