General Use Considerations

Transfection Tips

In our experience, MiniVectors will work with any plasmid transfection protocol. Cell types that are refractory to transfection with plasmid DNA are readily transfected with MiniVectors, thus expanding the opportunities for non-viral DNA delivery. Optimization of transfection protocols is recommended to gain the full benefits of our superior system.

We have had good results with a number of transfection reagents including, but not limited to, Lipofectamine™2000 (Life Technologies), FuGENE™(Promega), Xfect™(Clontech), Effectene™(Qiagen), and GenJet™(SignaGen). We recommend using the standard protocol for the reagent as a starting point, then optimize by varying the amount of DNA transfected. Because of its excellent transfection efficiency and small size (and consequently more copies per unit mass) you should require considerably less MiniVector and reagent.

Analysis of MiniVector Topology

It is important to note thatwhen analyzing topology for MiniVectors less than 1 kb, even high percentage agarose gels lack the resolution to separate supercoiled from relaxed MiniVector or linear DNA of the same size. We recommend polyacrylamide gel electrophoresis. Individual topoisomers may even be resolved and isolated using this technique. For optimum separation of topoisomers we recommend the addition of 10 mM CaCl2 to the running buffer. For more details of electrophoresis with MiniVector DNA, please see Fogg et al., J Phys (2006) or contact us at info@TwisterBiotech.com.

Protein-DNA interactions

MiniVector DNA is uniquely suited for studying proteins binding to supercoiled, physiologically relevant DNA substrate, without the complicating factors of a large excess of non-specific DNA sequence. The small size of MiniVectors means that unlike plasmids, it may be used in quantitative binding assays. We have had excellent results using MiniVector DNA in electrophoretic mobility shift assays (EMSAs).

Product Quality Control

Twister Biotech is committed to providing a high quality product. As part of our standard quality control (QC), we evaluate and document each batch of MiniVectors according to the following specifications:

1. Product will be clear and free of particulates based on visual inspection
2. DNA concentration will be calculated based on UV absorption at 260 nm
3. Product purity will be evaluated using agarose gel electrophoresis to confirm DNA band location at the expected molecular weight and absence of genomic or RNA contaminants
4. Product sheet contains a gel image of the purified MiniVector