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 fi rst 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.

15

191

309.5

30

137.5

320.5

60

71.5

294

Ethidium bromide/UV

CloneWell™/Safe Imager™

Exposure time (sec)

CFU

350

300

250

200

150

100

50

0

Get improved cloning effi cienciesExposure of your DNA sample to UV light during visualization may lead to DNA damage and reduced cloning effi ciencies. 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 effi ciency 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 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.

Select desired run protocol and press “Go” button

Retrieve DNA ready for cloningLoad sample

• 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

Gel-purify your DNA in 3 simple stepsE-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 purifi ed DNA band and you’re ready to clone. That’s it. No additional purifi cation 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.

E-Gel® CloneWell™ agarose gelsThe smartest way to gel-purify your DNA

View bands in real time and minimize DNA damageThe 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).

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For Research Use Only. Not for use in diagnostic procedures. © 2015 Thermo Fisher Scientifi c Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientifi c and its subsidiaries unless otherwise specifi ed. CO012548 0315

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.

2 4 8 16 2 4 8 16

NickedDNA

SupercoiledDNA

SYBR® Safe Stain and blue light exposure Ethidium bromide staining and UV exposure

Time (min)

Ordering information

Product Quantity Cat. No.

E-Gel® CloneWell™ Starter Kit 1 kit* G6500ST

E-Gel® CloneWell™ Agarose Gels with SYBR® Safe DNA Gel Stain 18 gels G6618-08

E-Gel® iBase™ and Safe Imager™ Real-Time Transilluminator Combo Kit 1 G6465

E-Gel® 96 High Range DNA Marker 100 applications 12352-019

E-Gel® Sample Loading Buffer 4 x 1.25 mL 10482-055

* Includes 1 E-Gel® iBase™ Power System and Safe Imager™ Real-Time Transilluminator, 1 E-Gel® 96 High Range DNA Marker, and 18 E-Gel® CloneWell™ Agarose Gels with SYBR® Safe DNA Gel Stain, 0.8% agarose.

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.

Retrieved bands

Samples loaded on CloneWell™ gel

1 2 Set 1 Set 2

4 kb3 kb2 kb1 kb

Retrieved bands

Samples loaded on CloneWell™ gel

1 2 Set 1 Set 2

4 kb3 kb2 kb1 kb

B

Retrieved bandsSamples loaded on CloneWell™ gel

Collection wells

Collect multiple DNA bands from the same gel laneWith 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 purifi cation is required (Figure 3).

A