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Invitrogen™ PureLink™ Fast Low-Endotoxin Midi Plasmid Purification Kit
Description
The simple precipitation-free protocol yields up to 0.4 mg plasmid, suitable for standard transfections and all molecular biology applications such as cloning and sequencing.
- Ultra-fast protocol—innovative silica-based columns enable purification of transfection grade plasmid DNA in under 30 minutes
- Low endotoxin plasmid—the resulting plasmid contains less than 1 EU/μg endotoxins
- High yields—isolate up to 0.4 mg of high-quality plasmid DNA
- Colored buffers—the kit features colored buffers that permit error-free visualization of complete bacterial cell lysis and subsequent neutralization
Specifications
Specifications
| Content And Storage | • Resuspension buffer, store at 4°C upon reciept • Lysis buffer, store at room temperature • Precipitation buffer, store at room temperature • Binding buffer, store at room temperature • Wash buffer 1, store at room temperature • Wash buffer 2 (concentrate), store at room temperature • Elution buffer, store at room temperature • Column assemblies, store at room temperature • Syringe filters, store at room temperature • Syringe plungers, store at room temperature • Collection tubes, store at room temperature |
| Format | Kit |
| Isolation Technology | Silica Column |
| Scale | Midi |
| For Use With (Application) | Cloning, PCR |
| No. of Reactions | 25 Preps |
| Prep Scale | Midi scale input culture of 25-100 mL to obtain yields of up to 200 μg |
| Product Line | PureLink |
| Product Type | Plasmid Purification Kit |
| Quantity | 25 Preps |
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Frequently Asked Questions (FAQs)
For any silica columns, elution with water is generally possible. However, a buffer is preferred for stability and accuracy of absorbance readings, as pure water can have a very low pH (4 - 5).
Endotoxins are typically any cell-associated bacterial toxins that are part of the outer surface of the cell wall of gram-negative bacteria. Endotoxins can influence cell growth, cell differentiation, contractility, and protein expression in mammalian cells. Endotoxins are released during bacterial lysis, and they can subsequently reduce transfection efficiency and protein expression levels. Please review the following article for more information about endotoxins: Butash KA et al. (2000) Reexamination of the effect of endotoxin on cell proliferation and transfection efficiency. Biotechniques 29(3): 610-614, 616, 618-619.
The ratio of absorbance at 260 nm to the absorbance at 280 nm (A260/A280) is typically used to measure purity of the sample. For DNA, the ideal A260/A280 ratio is 1.8, but it can be in the range of 1.7 - 1.9. The A260/A230 ratio is also used to determine if contamination is present. For DNA, the ideal A260/A230 ratio is between 1.8 and 2.0. DNA purity can also be examined by gel analysis. For plasmid DNA, look for a strong, single band (perhaps with a few extra bands representing multimers of the desired molecule). For genomic DNA, look for high average fragment sizes.
We offer the following silica column-based plasmid purification kits for isolation of endotoxin-free or low-endotoxin plasmid DNA:
- PureLink Expi Endotoxin-Free Maxi Plasmid Purification Kit (Cat. No. A31217, A31231, A33073) (<0.1 EU/µg endotoxins):
https://www.thermofisher.com/order/catalog/product/A31217
- PureLink Fast Low-Endotoxin Midi (Cat. No. A35892, A36227) or Maxi (Cat. No. A36228, A35895) Plasmid Purification Kits (<1 EU/µg endotoxins):
https://www.thermofisher.com/order/catalog/product/A35892
https://www.thermofisher.com/order/catalog/product/A36228
Our recommendation is to not exceed the maximum recommended culture volumes specified in the kit manuals; using larger culture volumes means increased cell mass, which will result in reduced plasmid yield and purity.
For Research Use Only. Not for use in diagnostic procedures.