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Tips from the Bench: Transfection Tip

Deliver microRNA (miRNA) Effectively

Transfect miRNAs In Vitro Using TransIT-X2® Dynamic Delivery System

TransIT-X2® Dynamic Delivery System is the reagent of choice for delivering miRNA, miRNA mimics or pre-miRNAs (see Figure 1). For miRNA transfection, follow the user protocol and simply substitute duplex miRNA for siRNA. TransIT-X2® delivers both plasmid DNA and/or miRNA efficiently and can be used for co-transfecting plasmid DNA and miRNA.

TransIT-TKO® and TransIT-siQUEST® Transfection Reagents can also effectively transfect duplex miRNAs into mammalian cells in culture. Ideally, all three reagents should be tested in parallel to determine the best solution for your application. Depending on the cell type, one reagent may have superior performance over the others. For cell-type specific recommendations, please consult the Reagent Agent® transfection database.

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Effective miRNA Delivery using TransIT-X2 Dynamic Delivery System Yields Decreased Levels of PTK9 mRNA

Effective miRNA Delivery using TransIT-X2® Dynamic Delivery System Yields Decreased Levels of PTK9 mRNA. TransIT-X2® Dynamic Delivery System and Lipofectamine® 2000 Transfection Reagent were used to transfect Pre-miR™ hsa-miR-1 miRNA Precursor or mirVana™ miRNA mimic, miR-1, both known to decrease PTK9 mRNA levels. A Pre-miR™ negative control was transfected to assess baseline mRNA levels. T47D cells were transfected in a 12-well plate using 3 µl of TransIT-X2® or Lipofectamine® 2000 and 50 nM miRNA according to each manufacturer's protocol. The amount of PTK9 mRNA was measured relative to 18s rRNA levels using qRT-PCR and then normalized to the mRNA levels of the negative control, 48 hours post-transfection. Error bars represent the standard deviation of triplicate wells.

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