| Technology | Silica-membrane and endotoxin-reduction technology |
| Format | Mini spin columns/96-well plates |
| Endotoxin content | <50 EU/μg DNA (EU = Endotoxin Units) |
| Sample material | Up to 5-ml bacterial culture |
| Vector size | <15 kbp |
| Typical yield | 5–20 μg |
| Elution volume | 30–200 μl |
| A260/A280 | 1.80–1.85 |
| Preparation time | 14 min/6 preps or 45 min/96 preps |
| Binding capacity | 35 μg |
Tech Note
NucleoSpin Plasmid Transfection-grade
Endotoxins are copurified during plasmid preparations from E. coli lysates and interfere with eukaryotic cell viability, making endotoxin reduction an essential step prior to cell transfection. NucleoSpin Plasmid Transfection-grade incorporates an innovative technology to reduce endotoxins from bacterial lysates compared to standard plasmid miniprep kits. The NucleoSpin Plasmid Transfection-grade and NucleoSpin 96 Plasmid Transfection-grade kits are designed for rapid, manual or automated, small-scale purification of transfection-grade plasmid DNA from E. coli.
If your cells are especially sensitive to endotoxin, see also Nucleobond Xtra EF products for the lowest possible endotoxin levels in your plasmid preps.
At-a-glance
Cell viability
Figure 1. pCMV-GFP plasmid (kindly provided by PlasmidFactory GmbH und Co. KG, Bielefeld, Germany) was purified from E. coli using NucleoSpin Plasmid Transfection-grade. Plasmids were transfected into HEK239 cells by lipofection (Lipofectamine 2000) or nucleofection (Lonza) with >90% transfection ratio in both cases. Cell survival was compared to controls without DNA addition. Similar ratios show that cell survival is not affected by DNA eluates purified with NucleoSpin Plasmid Transfection-grade.
Choose the right kit
Figure 2. Plasmids were purified from E. coli (DH5α) with different plasmid purification products from Macherey-Nagel and a quantitative chromogenic LAL-test was used to assess endotoxin content. As indicated, the endotoxin content strongly depended on the type of technology used for plasmid purification. Low endotoxin levels were detected after purification with NucleoSpin Plasmid Transfection-grade, resulting in a plasmid solution suitable for direct transfection of common cells.
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Tips for a successful transfection
This guide will help you select the right reagent and find useful tips for designing a successful transfection experiment. Tips include: how to plate your cells at the right density, how to make sure you are using plasmid DNA of sufficiently high quality, and how to assess transfection efficiency.
Transfection tips Plasmid transfection reagentsTakara Bio USA, Inc.
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