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This suggests that the same mechanism is at work to boost transfection efficiency for human stem cell lines.
hASCs and hMSCs have a similar phenotype [ 5, 12], so polymers that boost transfection efficiency of one cell type should produce a similar boost in the other.
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As observed in previous studies [ 10], the pluronic-block copolymer family shows the most consistent promise of boosting transfection efficiency.
Renilla luciferase plasmid, pRLnull (5 ng/well) was simultaneously transfected as an insertional control for transfection efficiency.
Transfection efficiency was determined at 24 h post-transfection.
Transfection efficiency was evaluated by RT-qPCR.
(J) Id3 siRNA transfection efficiency was found to be ~75%.
Cells were sorted for GFP 3 days post-transfection if transfection efficiency was below 90% (based on visual inspection).
Transfection efficiency was confirmed to be 30 45% using pHcRed1-Nuc Vector (Clontech) as a control.
The miRFP670 and miRFP670nano fluorescence signals were normalized to Rluc8 bioluminescence to account for transfection efficiency.
The activity was divided by the activity of Renilla control reporter to normalize the transfection efficiency.
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