optimize your transfections!€¦ · transfection reagent optimization poster author: mirus bio llc...
TRANSCRIPT
Optimize Your Transfections!
A1 Mix 12:1
A2 Mix 12:1
A3 Mix 23:1
A4 Mix 23:1
B1Mix 34:1
B2Mix 34:1
B3Mix 45:1
B4Mix 45:1
C1Cells Alone
Control
C2OptionalControl
C3OptionalControl
C4OptionalControl
5 tips to get the best results in your experiments
Get Your Ratios Right
It’s important to test multiple Reagent:DNA-ratios. Finding the optimal ratio for your cells will help unlock the highest transfection efficiency and decrease toxicity.
DNA-Reagent complex formation takes time. Allow your transfection reagent and DNA mixture to incubate for 15-30 minutes* at room temperature before adding the mix to your cells. Transfection efficiency will decrease if complex formation time exceeds one hour. *Incubation time varies by reagent, reference product protocol
Do: Use endotoxin-free, high quality DNA for all transfections. Contaminants like proteins, carbohydrates and lipids can negatively affect transfection efficiency and gene expression levels.
Don’t: Assume all cell types require the same dose of DNA. In some suspension cell lines, such as THP-1, transfecting higher amounts of DNA leads to higher transgene expression. Be sure to keep DNA:Reagent ratio constant when adjusting DNA dose.
15-30 min
The Complexity of Complexes
DNA Do’s and Don’ts
Divide and Conquer
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Use happy, healthy, and actively dividing cells to maximize your transfection efficiency. Mirus recommends plating your cells the night before transfection at a density that will promote cell division, and obtain 75-90% confluency at the time of transfection.