Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

Nanomagnetic Transfection
10,000 progenitor cells and 20,000 cardiomyocytes per well were seeded into 96 well tissue culture plates previously coated with 0.8 µg/µl rat tail collagen I, and onto extra cellular matrix coated tissue culture treated 96 well plates (Celprogen, California, USA) in 100 µl complete growth media and incubated for 24 h at 37°C with 5% CO2. Prior to transfection, 0.05 or 0.1 µg of pEGFP-N1 formed complexes with 0.2 or 0.35 µl NeuroMag (nanoTherics, Keele, UK) for 15 minutes in serum-free growth media containing antibiotics (Celprogen, California, USA) (Figure 1). NeuroMag, a MNP transfection vector was used for its non-interference with proliferation and low cytotoxicity, as observed during transfection of oligodendrocyte progenitor cells [19] and neural precursor/stem cells [21], [23]. After complex formation, 80 µl of complete medium was added, mixed and transferred, drop-wise, onto the appropriate well(s) containing prenatal human cardiac progenitor cells or adult cardiomyocytes. Plates were placed on a 96 well oscillating NdFeB (Rare earth magnet, an alloy of neodymium, iron and boron) magnet array following the magnefect nano II system protocol (Figure 1). Following 30 minutes exposure to the oscillating magnetic array, the culture plate was removed from the system and incubated at 37°C with 5% CO2 for 48 h. Lipofectamine 2000 (Invitrogen, Paisley, UK) and Nucleofector 2b (Lonza, Cologne, Germany) were evaluated for comparison and transfections were performed according to the manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Lipofectamine® 2000 Transfection Reagent below.

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Videos

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