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. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: Ther Deliv. 2013 Feb;4(2):267–277. doi: 10.4155/tde.12.155

Figure 4. Protamine-mediated precipitation of 3PO.

Figure 4

(A) 3PO was prepared by mixing 10 μg of DNA, 1 μg PBK10, and 10 μg protamine in a final volume of either 200, 100 or 50 μl. Mixtures were incubated at room temperature for 15 min, then spun in a microcentrifuge (15K RPM), while control samples were not. Supernatant absorbances were then measured at OD 260 to detect soluble DNA. (B) Increasing protamine excludes EtBr and forms precipitates. Plasmid DNA (0.35 μg, pEGFP) was incubated with indicated reagents for 15 min at room temperature, then electrophoresed on 0.8% agarose gel that was subsequently stained with EtBr to identify the DNA. (C) 3PO was prepared by mixing 0.35 μg PBK10, 3.5 μg DNA + increasing amounts of protamine in 100 μl final volume. After 15 min room temperature incubation, samples were spun in a micro-centrifuge (15K RPM), while control samples were not. Supernatant absorbances were then measured at 260 nm to detect soluble DNA.

EtBr: Ethidium bromide.