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E-Gel CloneWell II gels are compatible with the E-Gel Power Snap System
E-Gel CloneWell II 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 Invitrogen E-Gel Power Snap System, a compact, self-contained device with a built-in power supply and blue-light transilluminator, to run and visualize E-Gel CloneWell II agarose gels.
Figure 1. Three easy steps for separation and isolation of DNA bands with E-Gel CloneWell II 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 Power Snap System lets you capture bands of interest even if you miss the bands when they pass through the collection wells.
Exposure of your DNA sample to UV light during visualization may lead to DNA damage and reduced cloning efficiencies. Using E-Gel CloneWell II agarose gels with the E-Gel Power Snap System eliminates UV damage and improves cloning efficiency compared to conventional methods. Results obtained using the TOPO TA Cloning Kit after E-Gel CloneWell II gel purification, compared to a conventional method, are shown in Figure 2.
Figure 2. Improved cloning efficiency using E-Gel CloneWell II agarose gels with blue-light transillumination. 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 using blue-light transillumination. 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 sec. Both fragment samples were cloned using the TOPO TA Cloning Kit (Cat. No. K460040) and transformed into Invitrogen MultiShot TOP10 chemically competent cells (Cat. No. C40005). Shown are average numbers of colony-forming units (CFU) obtained for each exposure time and cloning method.
With E-Gel CloneWell II 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 II agarose gels, just retrieve the bands one at a time as they migrate into the collection well. No additional purification is required.
The E-Gel Power Snap System 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 3).
Figure 3. 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 defined 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.
E-Gel CloneWell II 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 Invitrogen E-Gel Power Snap System, a compact, self-contained device with a built-in power supply and blue-light transilluminator, to run and visualize E-Gel CloneWell II agarose gels.
Figure 1. Three easy steps for separation and isolation of DNA bands with E-Gel CloneWell II 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 Power Snap System lets you capture bands of interest even if you miss the bands when they pass through the collection wells.
Exposure of your DNA sample to UV light during visualization may lead to DNA damage and reduced cloning efficiencies. Using E-Gel CloneWell II agarose gels with the E-Gel Power Snap System eliminates UV damage and improves cloning efficiency compared to conventional methods. Results obtained using the TOPO TA Cloning Kit after E-Gel CloneWell II gel purification, compared to a conventional method, are shown in Figure 2.
Figure 2. Improved cloning efficiency using E-Gel CloneWell II agarose gels with blue-light transillumination. 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 using blue-light transillumination. 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 sec. Both fragment samples were cloned using the TOPO TA Cloning Kit (Cat. No. K460040) and transformed into Invitrogen MultiShot TOP10 chemically competent cells (Cat. No. C40005). Shown are average numbers of colony-forming units (CFU) obtained for each exposure time and cloning method.
With E-Gel CloneWell II 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 II agarose gels, just retrieve the bands one at a time as they migrate into the collection well. No additional purification is required.
The E-Gel Power Snap System 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 3).
Figure 3. 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 defined 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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For Research Use Only. Not for use in diagnostic procedures.
For Research Use Only. Not for use in diagnostic procedures.
For Research Use Only. Not for use in diagnostic procedures.