The smartest way to gel-purify your DNA

  • Gel-purify your DNA in 3 simple steps
  • Get improved cloning effi ciencies
  • View bands in real time and minimize DNA damage
  • Collect multiple DNA bands from the same gel lane

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E-Gel® CloneWell™ flyer

E-Gel® CloneWell™ gels are compatible with the E-Gel® iBase™ Power System

Watch the E-Gel® CloneWell™ Video

This video includes the step-by-step instruction of how to use E-Gel® CloneWell™ SYBR® Safe Gels.

Gel-purify your DNA in 3 simple steps

E-Gel® CloneWell™ agarose gels are double-comb gels with a twist. Load your sample into the top row and electrophorese until your band migrates into the bottom row (Figure 1). Then simply pipet out your purified DNA band and you’re ready to clone. That’s it. No additional purification kits or steps are required. Use the E-Gel® iBase™ Power System, a compact, self-contained device with a built-in power supply, and the E-Gel® Safe Imager™ Real-Time Transilluminator, to run and visualize E-Gel® CloneWell™ agarose gels.

Figure 1. Three easy steps for separation and isolation of DNA bands with E-Gel® CloneWell™ agarose gels. Samples are loaded into the top row of wells, bands separate during the gel run, and individual bands are collected from the bottom row as they enter those wells. Reverse-run functionality on the E-Gel® iBase™ Power System lets you capture bands of interest even if you miss the bands when they pass through the collection wells.

Get improved cloning effi ciencies

Exposure of your DNA sample to UV light during visualization may lead to DNA damage and reduced cloning efficiencies. Using E-Gel® CloneWell™ agarose gels with the E-Gel® iBase™ Power System and Safe Imager™ Real-Time Transilluminator (Cat. No. G6465) eliminates UV damage and improves cloning efficiency compared to conventional methods. Results obtained using the TOPO® TA Cloning® Kit after E-Gel® CloneWell™ gel purifi cation, compared to a conventional method, are shown in Figure 2.

Figure 2. Improved cloning effi ciency using E-Gel® CloneWell™ agarose gels with the E-Gel® iBase™ Power System and Safe Imager™ Real-Time Transilluminator. A PCR reaction containing an 850 bp amplicon was separated on either an E-Gel® CloneWell™ agarose gel or a traditional agarose gel containing ethidium bromide. The DNA band retrieved from the E-Gel® CloneWell™ agarose gel was visualized on the E-Gel® Safe Imager™ Real-Time Transilluminator. DNA separated on the traditional gel was viewed using UV light and isolated by first cutting a gel slice with a razor blade and then using a commercially available gel extraction kit. In both cases, the exposure of the DNA to the light source was 15, 30, or 60 seconds. Both fragment samples were cloned using the TOPO® TA Cloning® Kit (Cat. No. K4600-40) and transformed into TOP10 chemically competent cells (Cat. No. C400-05). Shown are average numbers of colony forming units (CFU) obtained for each exposure time and cloning method.

Collect multiple DNA bands from the same gel lane

With E-Gel® CloneWell™ agarose gels, you can retrieve multiple DNA bands from the same sample/lane, thus saving materials and time. Avoid the hassle of cutting out gel bands and using multiple columns for further gel extraction. With E-Gel® CloneWell™ agarose gels, just retrieve the bands one at a time as they migrate into the collection well. No additional purification is required (Figure 3).

Figure 3. Collect multiple DNA bands from the same gel lane. (A) Samples of 10 µL and 5 µL of the High DNA Mass Ladder (Cat. No. 10496-016) were loaded onto an E-Gel® CloneWell™ agarose gel in lanes 1 and 2, respectively. The four smallest bands were isolated by pipetting them out after each had migrated into the collection well. (B) Each of the retrieved bands was run in a separate lane on a 1.2% E-Gel® agarose gel (Cat. No. G5218-01). Set 1 was isolated from lane 1 in gel A. Set 2 was isolated from lane 2 in gel A.

View bands in real time and minimize DNA damage

The E-Gel® iBase™ and Safe Imager™ Real-Time Transilluminator combination allows real-time viewing of DNA band migration through E-Gel® agarose gels containing SYBR® Safe DNA Gel Stain. Unlike UV light, the blue light used in this system causes minimal DNA damage (Figure 4).

Figure 4. Use of SYBR® Safe DNA Gel Stain and a blue-light transilluminator causes minimal DNA damage. Equivalent fractions of supercoiled DNA stained with SYBR® Safe DNA Gel Stain or ethidium bromide were exposed to blue light (Safe Imager™ 2.0 Blue-Light Transilluminator) or UV light, respectively, for defi ned periods of time and evaluated by agarose gel electrophoresis. A slower-migrating species is indicative of a linear or relaxed circular vector that results from DNA nicking or strand breaks.

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