ChargeSwitch — Pro Plasmid Miniprep Kit

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Researchers have been performing plasmid minipreps for years—but they could be using an easier, faster, safer method and getting better results. Today, the majority of the plasmid miniprep methods in use require a silica membrane for DNA binding and purification. While generally effective, this technology suffers from a number of limitations, including the possible introduction of contaminants into downstream applications, compromising your results (Table 1). Other methods employ anion exchange to purify plasmid DNA. This method is effective but is expensive, slow, and requires a precipitation step during which you may lose sample. But, there is a better way—ChargeSwitch® pH-dependent technology. Now you can purify plasmids without introducing inhibitors such as ethanol and chaotropic salts and obtain a cleaner purified plasmid for optimal downstream performance in PCR, sequencing, and transformation experiments.

Table 1. ChargeSwitch Pro Plasmid Miniprep Kits overcome the limitations of traditional plasmid purification

The ChargeSwitch protocol retains the ease and familiarity of common silica spin-column protocols with binding, washing, and elution steps (Figure 1, Table 2), making adapting to the ChargeSwitch method effortless.

Just add your sample to the Plasmid Miniprep Kits

The ChargeSwitch-Pro Plasmid Miniprep Kit provides all reagents and components required to process your sample. There is no need to add ethanol to any buffers or search for additional collection tubes. All you need to supply is your E. coli culture.

Table 2. ChargeSwitch Pro Plasmid Miniprep Kits specifications.

Since ChargeSwitch Technology does not require the use of ethanol, chaotropic salts, or organic solvents for nucleic acid purification, the resulting DNA is free of potentially inhibitory compounds, improving DNA integrity for better downstream PCR performance (Figure 2). Expect to see improved cloning due to higher transformation efficiencies (Figure 3), faster restriction enzyme digestions (Figure 4), sequencing reads in excess of 800 base pairs with Phred scores >20, and a higher degree of supercoiling (Figure 5).

Figure 2.  ChargeSwitch-Pro reagents do not contain inhibitory components that may interfere in downstream PCR reactions. The inhibitory reagents used in most plasmid purification techniques are difficult to detect when measuring DNA quality on gels or by UV spectrophotometry. They can, however, inhibit PCR. The action gene was amplified by PCR from human placental DNA, in a 50-µL reaction spiked with varying volumes of ethanol, isopropanol (IPA), competitor wash buffers, guanidinium isothiocyanate (GTC) and ChargeSwitch wash buffer. Ten microliters of amplified product run on a 1% agarose gel and stained with ethidium bromide.

Figure 3. Higher transformation efficiencies with plasmid purified using the ChargeSwitch-Pro Kit. The ChargeSwitch-Pro Plasmid Miniprep Kit and a competitor silica prep kit (Q) were used to purify pUC19 plasmid DNA from 5 mL of LB culture. One Shot® TOP10 Chemically Competent E. Coli was transformed using 10 pg of the purified pUC19 plasmid DNA, or OneShot TOP10 Electrocomp E. Coli was transformed with 5 pg of the purified pUC19 plasmid DNA. pUC19 plasmid DNA supplied with the competent cells was used as a control. Error bars represent 1 SE (n=12).

Figure 4.  Improved restriction enzyme digestion kinetics with plasmid purified using the ChargeSwitch-Pro Kit. ChargeSwitch-Pro Miniprep column-purified DNA (C) can be digested faster than DNA purified using a silica column from a competitive kit (Q), pUC19 (A) or pLadder (B) purified with the ChargeSwitch-Pro kit were completely digested in about 30 min, whereas the silica-purified sample was only partly digested after 90 min.

注）精製バッファーに対しPCR反応液が、競合キットプロトコルでは3：1なのに対し、ChargeSwitch-Pro PCR Clean-up カラムでは1：1にできるので、 大容量のPCRや酵素の反応液を最大500μlまで1回のスピンで処理できます。