ASO transfection of iPSC-derived cells
James Evans
Abstract
A protocol for transfecting iPSC-derived midbrain dopaminergic neurons with antisense oligonucleotides (ASOs).
Steps
ASO transfection of iPSC-derived mDA neurons
Make up transfection mix in N2B27. The transfection mix should be 1/5 of the final volume in the well.
Transfection mix should have:
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0.48 % DharmaFECT DharmaFECT transfection reagent
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ASO (adjust concentration depending on cell type, ASO chemical modification, and knockdown required). Calculate final concentration required in the well not in the transfection mix.
Example - 1000 ul final volume in the well with 300 nm of ASO = 200 ul of N2B27 + 0.96 ul of DharmaFect + 3 ul of 100 uM ASO Stock
Vortex transfection mix and leave at room temperature for 30 minutes.
Aspirate media from cells and replace with the transfection mix. After adding the transfection mix add the rest of the media to the cells to make up the final volume.
