Quantifying SCN2A Expression in iNeurons

RNA from iNeurons cultured were extracted using the RNeasy Mini kit (Qiagen). cDNA was generated using the qScript cDNA SuperMix (QuantaBio). ddPCR was performed as previously described (Deneault et al., 2018 (link)). Quantification was normalized to the control of each condition and run along a no template control. The data was analyzed and produced on the QuantaSoft software (Bio-Rad). The following assays were used: SCN2A exon 4–5 (Hs01109883_m1, ThermoFisher) and TBP (Hs00427620_m1, ThermoFisher).

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Brown C.O., Uy J.A., Murtaza N., Rosa E., Alfonso A., Dave B.M., Kilpatrick S., Cheng A.A., White S.H., Scherer S.W, & Singh K.K. (2024). Disruption of the autism-associated gene SCN2A alters synaptic development and neuronal signaling in patient iPSC-glutamatergic neurons. Frontiers in Cellular Neuroscience, 17, 1239069.

Publication 2023

RNeasy Mini kit qScript cDNA SuperMix QuantaSoft software Hs00427620_m1

Corresponding Organization : University of Toronto

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Variable analysis

independent variables

RNA extraction method (RNeasy Mini kit, Qiagen)
CDNA generation method (qScript cDNA SuperMix, QuantaBio)
DdPCR method (as previously described in Deneault et al., 2018)

dependent variables

Quantification of gene expression for SCN2A exon 4-5 and TBP

control variables

Normalization of quantification to the control of each condition
Inclusion of a no template control

controls

Positive control: Quantification normalized to the control of each condition
Negative control: No template control

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