Source: Lo, J. C. W. et al., Efficient and Cost Effective Electroporation Method to Study Primary Cilium-Dependent Signaling Pathways in the Granule Cell Precursor. J. Vis. Exp. (2021)
This video demonstrates the electroporation technique that introduces fluorescent protein-encoding plasmid DNA into mouse primary cerebellar granule cell precursors.
1. Electroporation for Hh receptor transgene overexpression: pEGFP-Smo
Poring Pulse Setting | Transfer Pulse Setting | |||
Mouse Primary GCPs | Primary neurons | Mouse Primary GCPs | Primary neurons | |
Voltage | 275 V | 275 V | 20 V | 20 V |
Length | 1 ms | 0.5 ms | 50 ms | 50 ms |
Interval | 50 ms | 50 ms | 50 ms | 50 ms |
No. | 2 | 2 | 5 | 5 |
D rate | 10% | 10% | 40% | 40% |
Polarity | + | + | ± | ± |
Table 1: The electroporation parameters of mouse primary GCPs and primary neurons using Super Electroporator NEPA21 TYPE
The authors have nothing to disclose.
B27 supplement | Life Technologies LTD | 17504044 | |
GlutamMAXTM-I ,100x | Gibco, Life Technologies | 35050061 | L-glutamine substitute |
Matrigel | BD Biosciences | 354277 | Basement membrane matrix |
Neurobasal | Gibco, Life Technologies | 21103049 | |
Poly-D-lysine Hydrobromide | Sigma Aldrich | P6407 | |
CU 500 cuvette chamber | Nepagene | CU500 | |
EPA Electroporation cuvette (2 mm gap) | Nepagene | EC-002 | |
Opti-MEM | Life Technologies LTD | 31985070 | Reduced-serum medium for transfection |
pEGFP-mSmo | Addgene | 25395 | |
Super Electroporator NEPA21 TYPE II In Vitro and In Vivo Electroporation | Nepagene | NEPA21 | Electroporator |