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查看更多产品信息 E-Gel™ Agarose Gels with SYBR™ Safe DNA Gel Stain, 2% - FAQs (G521802)
67 个常见问题解答
与溴化乙锭相似，SYBR Safe DNA凝胶染料与DNA的迁移方向相反。由于只有凝胶的最底部染料浓度会在电泳过程中降低，所以使用SYBR Safe DNA凝胶染料制备的凝胶没有实际影响。在电泳结束后用SYBR Safe DNA凝胶染料染胶，可部分抵消这种作用。染料溶液不应该加入到电泳缓冲液中，因为这可能会导致染料在电极上分解，并将有毒的挥发性化合物释放到空气中。
SYBR染料染色的琼脂糖胶可以用以下仪器进行观察：标准的UV紫外透射仪，应配置有紫外范围或者介于470和530 nm之间激发光波长的激光扫描仪；或者蓝光透射仪。
这里是一些建议：
•用酒精和Kimwipes纸巾擦拭胶盒。
•对相机镜头进行清洁。
•对成像系统的曝光时间和亮度选项进行调整。
请确保您没有过量上样并且在移除梳子时胶孔没有被损坏。
虽然我们建议将它们保存在室温，但这些凝胶仍然可以使用。在电泳前将这些凝胶平衡至室温以获得最佳效果。
首先，检查你的琼脂糖凝胶的浓度。较高浓度的凝胶能帮助你分离较小分子量的分子，而较低百分比的凝胶能帮你分离较大分子量的分子。
上样缓冲液是可选的。如果没有使用缓冲液，样品可直接加到孔内，或者你可以使用去离子水或者TE缓冲液稀释样本。如果你想使用上样缓冲液，请参考以下配方：<
E-Gel 琼脂糖凝胶（包含EX）
10 mM Tris-HCl, pH 7.5
1 mM EDTA
0.005% 溴芬蓝
0.005% 二甲苯青 FF
E-Gel CloneWell和E-Gel SizeSelect凝胶
10 mM Tris-HCl, pH 7.5
1 mM EDTA
此外，您可以使用 10X BlueJuice上样缓冲液或 TrackIt上样缓冲液。对此缓冲液进行50倍到200倍稀释以获得最佳E-Gel琼脂糖凝胶电泳结果。
每个泳道或者每个条带推荐的DNA上样量为20–100 ng。对于大部分E-Gel琼脂糖凝胶来说每个条带DNA上样量最多为500ng，而对于使用SYBR Safe染料的E-Gel凝胶，每条带的DNA的上样量最多为700ng。
您可以使用我们的E-Editor软件，完成电泳后它能帮您调整图象。E-Editor 2.0软件只适用于PC操作系统，而更早的E-Gel 96编辑软件仍适用于 Mac操作系统并能对 E-Gel 96和E-PAGE 96凝胶电泳图进行调整。不过原来的软件不适用于E-Gel 48或E-PAGE 48凝胶。请前往www.thermofisher.com在搜索框输入“E-Editor software”下载E-Editor软件。您可以使用E-Gel 成像系统进行数据分析。
高通量E-Gel琼脂糖凝胶有交错的胶孔，而且其电泳是基于中性pH值的内部缓冲系统，而非基于离子交换基质。高通量E-Gel琼脂糖凝胶不能被打开，而且要使用E-Gel E-Base系统而不是iBase或PowerBase系统进行电泳。
没有，我们测试过在E-Gel琼脂糖凝胶所有上样道加水及仅在样本道加水，结果并未观测到明显的差异。但是对于E-Gel CloneWell琼脂糖凝胶和E-Gel SizeSelect琼脂糖凝胶，根据使用手册加水则很重要。
E-Gel EX琼脂糖凝胶可用于分离DNA或RNA。可以在非变性或变性条件下进行RNA的分离。请注意我们没有对凝胶是否含有RNA酶进行过质控检测。下面是分别针对非变性或变性样本进行电泳时的建议：
非变性条件
1. 将RNA样本与15 µL不含RNA酶的水混合。
2. 不要加热。将全部样本上样到E-Gel EX琼脂糖凝胶上。
3. 电泳30分钟。
变性条件*
1. 将15 µL RNA上样缓冲液与 1–5 µL RNA（1–5 µg）混合。
2. 将样本在65°C加热10分钟，对RNA进行变性。
3. 加热后立刻将样本置于冰上。
4. 将全部样本上样到E-Gel EX琼脂糖凝胶上。
5. 电泳30分钟。
*唯一与E-Gel EX系统兼容的变性剂是甲酰胺，浓度在50–95%。65℃加热5分钟足以将样品变性。 E-Gel EX琼脂糖凝胶电泳时，使用其它的变性剂，如乙二醇、甲醛或尿素会导致分离效率极低，且带形极差。此外，我们不建议将在RNA上样缓冲液中制备的样品与在水中制备的样品在同一凝胶上电泳。
对于含有SYBR及溴化乙啶的E-Gel琼脂糖凝胶胶盒，我们推荐使用E-Gel Opener（货号G530001）打开凝胶盒。E-Gel EX琼脂糖凝胶、 E-Gel SizeSelect 琼脂糖凝胶和 E-Gel GO!琼脂糖凝胶可以用凝胶切刀 （EI9010）打开。我们不推荐打开E-Gel 48琼脂糖凝胶胶盒或者E-Gel 96琼脂糖凝胶胶盒。
为了创造无缓冲液系统，每个E-Gel盒内的凝胶与正负电极间包含两种独立的离子交换基质。离子交换基质提供缓冲离子库，为整个凝胶不断提供乙酸、Tris及乙啶离子。这项专利技术产生了一个持续的具有高缓冲能力的电场。E-Gel 凝胶使您无需直接暴露于溴化乙啶，并无需配制及废弃液体缓冲液，从而节约时间并防止浪费。
E-Gel系统的E-Gel SYBR及溴化乙啶凝胶是使用离子交换基质的无缓冲液TAE系统预制凝胶。凝胶本身由一个UV半透过盒包围。
完整RNA的28S:18S条带亮度比例应为2:1。你可能会在0.5 kb至9kb区间看到模糊的RNA拖尾条带，这表明样本中含有mRNA。请点击此处(https://www.thermofisher.com/cn/zh/home/references/ambion-tech-support/rna-isolation/tech-notes/assessing-rna-quality.html)查看图片或了解更多评估RNA质量的信息。
对于非变性RNA电泳，我们推荐使用 E-Gel 预制胶电泳系统。请注意E-Gel 琼脂糖凝胶并不保证完全不含RNA酶。但是，我们许多用户经常使用E-Gels琼脂糖凝胶成功的完成了RNA分析。如果使用E-Gel琼脂糖凝胶进行RNA电泳，可以使用任何适用于非变性DNA电泳的上样缓冲液。
对于变性RNA电泳，可以从以下变性试剂中进行选择，包括甲醛、乙二醛、甲酰胺、甲基汞。变性条件会破坏氢键，因此RNA在电泳时没有二级结构，作为单链分子而迁移。
对变性RNA电泳，我们的E-Gel EX琼脂糖凝胶可以使用。唯一与E-Gel EX系统兼容的变性剂是甲酰胺，50-95%。使用其他变性剂会导致条带分离较差，带型也不理想。请注意，我们不建议将在RNA上样缓冲液中制备的样品与在水中制备的样品在同一凝胶上电泳。RNA上样缓冲液配方及变性电泳条件如下：
RNA上样缓冲液：
去离子化甲酰胺：200 μL
10X MOPS-EDTA-乙酸钠缓冲液（0.4 M MOPS, pH 7.0，0.1 M乙酸钠，10 mM EDTA)：40 μL
去离子化甲醛：76 μL
水：14 μL < br / > < br / >
变性电泳条件：< br / >
1. 将15 μL RNA上样缓冲液与1-5 μL RNA (1-5 μg)混合。< br / >
2. 在65摄氏度下加热10分钟使RNA变性。< br / >
3. 加热后立即将样品放在冰上。< br / >
4. 将全部样品加载到E-Gel EX琼脂糖凝胶上。
5. 电泳30分钟。
对于不用甲醛作为变性剂的RNA变性电泳，我们推荐使用 NorthernMax-Gly 试剂盒（货号AM1946）。使用此试剂盒时，RNA样本在乙二醛／DMSO上样缓冲夜中变性，然后在含有乙二醛的琼脂糖凝胶中进行电泳。
琼脂糖因为无毒、易用且分离范围大而被广泛使用。我们提供预制E-Gel琼脂糖凝胶，也提供试剂用于配制琼脂糖凝胶。聚丙烯酰胺凝胶通常用于对10–3,000bp大小的DNA分子进行高分辨率分析。我们提供预制Invitrogen TBE聚丙烯酰胺凝胶和UltraPure试剂。
Similarly to ethidium bromide, SYBR Safe DNA Gel Stain runs in the opposite direction of the migrating DNA. This has no practical effect on the use of gels cast with SYBR Safe DNA Gel Stain, as only the very bottom of the gel will have a lower concentration of stain. This effect can be partially counteracted by staining the gel with SYBR Safe DNA Gel Stain after electrophoresis. Solutions of dye should not be added to the running buffer as this can cause breakdown of the dye at the electrodes and release toxic volatile compounds into the air.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
SYBR Safe-stained gels can be visualized with a standard UV transilluminator, a laser-based scanner equipped with an excitation source in the UV range or between 470 and 530 nm, or a blue-light transilluminator.
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Here are some suggestions:
- Try cleaning the cassettes with alcohol and Kimwipes wipers.
- Try cleaning the camera lens.
- Try to adjust the exposure time and brightness options of the documentation system you are using.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Please ensure that you have not overloaded the well and that the wells were not damaged during comb removal.
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While we recommend storage at room temperature, these gels will still be usable. Bring the gels to room temperature prior to the run for optimal conditions.
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First check the percentage of your agarose gel. A higher percentage will help you to resolve smaller molecular weights while a lower percentage will help you to resolve larger molecular weights.
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Loading buffer is optional. Samples can be loaded directly into the wells if no buffer is used, or you can dilute them with deionized water or TE buffer. If you want to use a loading buffer, please see the recipes below:
E-Gel agarose gels (including EX)
10 mM Tris-HCl, pH 7.5
1 mM EDTA
0.005% bromophenol blue
0.005% xylene cyanol FF
E-Gel CloneWell II and E-Gel SizeSelect II agarose gels
10 mM Tris-HCl, pH 7.5
1 mM EDTA
Alternatively, you can use 10X BlueJuice Gel Loading Buffer or TrackIt Loading Buffer. Dilute this buffer 50- to 200-fold to obtain optimal results with E-Gel agarose gels.
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To determine the amount of DNA to load per well for your specific E-Gel agarose gel, please refer to the table on page 14 of the E-Gel Power Snap Electrophoresis System user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017050_egel_powersnapsystem_UG.pdf).
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We offer our E-Editor Software, which can help you align images after a gel run. The E-Editor 2.0 Software is only available for PCs, but the older E-Gel 96 Editor software is still available for the Mac operating system and can align images from E-Gel 96 and E-PAGE 96 agarose gels. However, the original software is not compatible with E-Gel 48 or E-PAGE 48 agarose gels. Please go to www.thermofisher.com and enter "E-Editor software" in the main search to download the E-Editor Software. You can use the E-Gel Imager System for data analysis.
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p>High-throughput E-Gel agarose gels have staggered wells, and are based on a neutral-pH internal buffer system as opposed to an ion exchange matrix. High-throughput E-Gel agarose gels cannot be opened, and should be run on the E-Gel E-Base system.
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If you do not have enough samples to use all the wells, fill each empty well with the same volume of water as the loaded samples. For E-Gel CloneWell II agarose gels and E-Gel SizeSelect II agarose gels, it is important to add the water according to the respective manual.
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Our E-Gel agarose gels can be used for either DNA or RNA separation. RNA separation occurs under non-denaturing or denaturing conditions. Please note, our gels are not QC tested for the presence of RNases. See our suggestions below for running your non-denaturing or denaturing samples:
Non-denaturing conditions
1. Mix RNA sample with 15 µL of RNase-free water.
2. Do not heat. Load the entire sample onto the E-Gel agarose gel.
3. Electrophorese for 30 min.
Denaturing conditions
1. Mix 15 µL of RNA loading buffer with 1-5 µL of RNA (1-5 µg).
2. Heat samples at 65 degrees C for 10 min to denature RNA.
3. Place samples on ice immediately after heating.
4. Load entire sample onto E-Gel agarose gel.
5. Electrophorese for 30 min.
Note:
- The only denaturing agent that is compatible with the E-Gel system is formamide, 50-95%. Heating the sample for 5 min at 65 degrees C should be sufficient for denaturing. Using other denaturing agents like glyoxal, formaldehyde, or urea will result in very poor separation and band morphology on E-Gel agarose gels.
- Additionally, we do not recommend running samples with RNA loading buffer on the same gel as samples loaded with water.
- E-Gel EX Double Comb agarose gels and E-Gel Double Comb agarose gels with SYBR Safe stain have not been tested for use in RNA applications.
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We recommend using the E-Gel Opener (Cat. No. G530001) for the SYBR and ethidium bromide E-Gel agarose gels. The E-Gel EX agarose gels, E-Gel SizeSelect agarose gels, and E-Gel GO! agarose gels can be opened with the Gel Knife (Cat. No. EI9010). We do not recommend opening the E-Gel 48 agarose gels or the E-Gel 96 agarose gels.
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To create a bufferless system, each E-Gel cassette contains two unique ion exchange matrices that lie between the running gel and the electrodes. The ion exchange matrices provide a buffer-ion reservoir that supplies a continuous flow of Tris, acetate, and dye ions throughout the gel. This patented technology results in a sustained electric field with enhanced buffering capacity. E-Gel gels thus eliminate the need for you to prepare and dispose of liquid buffer, thus saving time and waste.
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The E-Gel agarose gel system is a precast bufferless TAE system that uses ion exchange matrices. The gels themselves are enclosed within a semi-UV-transparent cassette. This patented technology results in a sustained electric field with enhanced buffering capacity. E-Gel gels thus eliminate the need for you to prepare and dispose of liquid buffer, thus saving time and waste.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Intact RNA should have a 2:1 ratio of 28S:18S bands. You may see a smear of RNA that extends from <9 kb to 0.5 kb, indicating the presence of mRNA in the sample. To see an image or to read more about RNA assessment, visit this website (https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/agarose-gel-electrophoresis-of-rna.html).
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For nondenaturing RNA electrophoresis, we recommend using our E-Gel Precast Agarose Gels. Please note that E-Gel Agarose Gels are not validated to be RNAse-free. However, many of our customers routinely use E-Gel Agarose Gels for RNA analysis with success. If RNA is run on an E-Gel Agarose Gel, any loading buffer that would be used for nondenaturing RNA electrophoresis should be fine.
For denaturing RNA electrophoresis, there are several denaturing agents to choose from, including formaldehyde, glyoxal, formamide, and methyl mercury. Denaturing conditions disrupt hydrogen bonding so that RNA runs without secondary structure, as single-stranded molecules.
For denaturing RNA electrophoresis, our E-Gel EX Agarose Gels can be used. The only denaturing agent that is compatible with the E-Gel EX system is formamide, 50-90%. Using other denaturing agents will result in poor band separation and morphology. Please note that we do not recommend running samples prepared in RNA loading buffer on the same gel with samples prepared in water. Please see below for the RNA loading buffer recipe and denaturing electrophoresis conditions:
RNA Loading Buffer:
Deionized formamide: 200 µL
10X MOPS-EDTA-Sodium Acetate Buffer (0.4 M MOPS, pH 7.0, 0.1 M sodium acetate, 10 mM EDTA): 40 µL
Deionized formaldehyde: 76 µL
Water: 14 µL
Denaturing Electrophoresis Conditions:
1. Mix 15 µL of RNA loading buffer with 1-5 µL of RNA (1-5 µg).
2. Heat samples at 65 degrees C for 10 min to denature RNA.
3. Place samples on ice immediately after heating.
4. Load entire sample onto an E-Gel EX agarose gel.
5. Electrophorese for 30 minutes.
For denaturing RNA electrophoresis under formaldehyde-free conditions, we recommend using our NorthernMax-Gly Kit (Cat. No. AM1946). With this kit, RNA samples are denatured in glyoxal/DMSO loading buffer and run on a glyoxal-containing agarose gel.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Agarose is commonly used as it is nontoxic, easy to use, and offers a broad range of separation. We offer precast E-Gel Agarose Gels or reagents to pour your own agarose gels. Polyacrylamide gels are typically used for high resolution of DNA molecules that range in size from 10-3,000 bp. We offer precast Invitrogen TBE polyacrylamide gels and UltraPure reagents.
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Yes, SYBR Safe stain is easily removed from nucleic acids by ethanol precipitation or by the ethanol wash steps used for purification spin columns.
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Several E-Gel products are available with SYBR Safe DNA gel stain. These gels can be used in the same manner as their ethidium bromide counterparts, with the additional safety and application benefits of SYBR Safe. To learn more about these products, search "E-Gel Precast Agarose Gels" from the Thermo Fisher Scientific website home page.
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We have found a distinct advantage to using SYBR Safe stain rather than ethidium bromide when purifying DNA from gels for downstream use. SYBR Safe stain is compatible with blue light imaging systems as well as UV. Using blue light to visualize the DNA allows you to purify a band with virtually no UV-induced nicking or crosslinking. This can dramatically increase cloning efficiency. Data from one such experiment showing higher cloning efficiency with PCR products visualized with SYBR Safe and blue light vs. ethidium bromide and UV light can be seen on the information page for Safe Imager 2.0 Blue-Light Transilluminator (Cat. No. G6600) and on the SYBR Safe home page.
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We recommend that SYBR Safe stain be protected from light during storage and gel staining. However, it is sufficiently stable to withstand UV illumination for >30 minutes; realistically, hours of constant UV or bright room light exposure are required to cause any significant loss of signal.
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SYBR Safe stain yields the same sensitivity as ethidium bromide - roughly 500 pg/band in a minigel for fragments larger than 200 bp viewed on a 300 nm transilluminator.
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Many whitening agents used in clothing, some synthetic fibers, as well as some fungi and bacteria, fluoresce at the same wavelength as SYBR Safe stain. These contaminants within or on the surface of the gel may produce this “speckling”. This can be avoided by being careful with preparation of the gel (i.e. try to keep the gel dust free). Alternatively, to obtain a publication quality image you may be able to preferentially photobleach some contaminants by leaving the gel on the UV box for 15-30 minutes, where the speckles will disappear before the SYBR stain photobleaches.
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SYBR Safe DNA gel stain has two main excitation peaks: in the UV region at 280 nm, and in the visible region at 502 nm. Thus, 254 nm or 300 nm UV excitation will work, as will 488 nm lasers, 470 nm LEDs, and broad blue excitation (such as the Safe Imager 2.0 Blue-Light Transilluminator, Cat. No. G6600). Maximal excitation occurs at 502 nm; the Safe Imager 2.0 Blue-Light Transilluminator is therefore the best choice for excitation of SYBR Safe DNA gel stain. The full excitation and emission spectra for SYBR Safe DNA gel stain are provided online and can also be found in the protocol.
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Some ethidium bromide filters allow the transmission of all light above 500 nm. These filters (which are often yellow in color) and their associated camera settings can be used with SYBR Safe DNA gel stain, usually with only minor adjustments to the exposure or gain. Other ethidium bromide filters (often red in color) only transmit light around or above 600 nm; these filters and their associated camera settings are not suitable for use with SYBR Safe DNA gel stain.
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Stained gels can be photographed using Polaroid 667 black-and-white print film and SYBR Safe photographic filter (Cat. No. S37100). Invitrogen SYPRO photographic filter (Cat. No. S6656) or a Kodak Wratten #9 filter also work well. Table 5 in the SYBR Safe product manual lists a filter selection guide for different instruments.
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Please see the SYBR Safe home page for a list of recommended filters and settings for several different gel documentation instruments. You can find it by searching "SYBR Safe DNA Gel Stain" from the Thermo Fisher Scientific website home page.
If your system is not listed, please contact the instrument manufacturer for a recommendation. Note that the excitation and emission spectra of SYBR Safe gel stain are very similar to those of SYBR Green I, SYBR Green II, and SYBR Gold gel stains, as well as fluorescein (FITC). Therefore, filters appropriate for these dyes can also be used.
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The SYBR Safe photographic filter (Cat. No. S37100) is a Wratten #9 gelatin filter. This filter is a 75 mm x 75 mm sheet of plastic that should be mounted in front of the lens of the camera. With a Polaroid camera and B&W film (#667), the filter may be taped inside the hood or mounted in a cassette and snapped in place inside the hood (the opening in which the camera lens is mounted upon).
For other camera systems, this sheet may be mounted in a cassette or filter-housing and placed in front of the camera lens. An alternative is to use thread-on glass filters of the same rating available from most camera supply vendors. Please note - not all camera systems require use of the SYBR Safe filter. See the SYBR Safe home page for a list of recommended filters and settings for several different instruments - you can find the page by searching "SYBR Safe DNA Gel Stain" from theThermo Fisher Scientific website home page.
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DNA bands stained with SYBR Safe stain may be visible by eye on a 300 nm transilluminator if there is a sufficient amount of DNA per band. However, optimal detection is obtained by photographing the gel using a Wratten #9 emission filter (or other filter with a similar rating) with your CCD or film camera. With UV transilluminators (light boxes), UV bulbs may also emit some infrared (IR) wavelengths; if your camera lens is not specially coated to block IR, an IR-blocking filter is needed to prevent the appearance of the UV bulbs under your gel.
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Some institutions and municipalities have approved the disposal of SYBR Safe DNA Gel Stain directly into their waste water systems and regular trash receptacles. However, disposal regulations vary; please contact your safety office or local municipality for disposal guidelines. For more information on environmental testing and safety standards for SYBR Safe stain, please visit our main SYBR Safe informational page, which you can find by searching "SYBR Safe DNA Gel Stain" from the Thermo Fisher Scientific website home page.
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Migration speed through gels is correlated to the compactness of the DNA molecule. Circular DNA can exist as a relaxed circle (if one of the strands is nicked), and this form migrates very slowly. If the DNA circle is supercoiled, it adopts a more compact form and will migrate much more quickly. If both strands of the DNA circle are cleaved and the molecule is linear, that form will also migrate more quickly than the relaxed circle, but not as quickly as the supercoiled circle.
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Pre-run is no longer necessary for any of our E-Gel agarose gels.
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E-Gel agarose gels are routinely used in-house in R&D and manufacturing for RNA analysis with excellent results. However, the gels are not guaranteed to be “RNAse-free”. The manufacturing process is designed to avoid contamination of any type, but not RNAses specifically.
If you do want to try it, any loading buffer that would be used for non-denaturing RNA electrophoresis should be fine for E-Gel agarose gels. Depending on your application, these gels may or may not be suitable for use.
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Do not run E-Gel agarose gels longer than 40 min for the single comb gels or longer than 20 min for the double comb gels as longer run times will cause ions to get depleted and will damage the gel. Do not run E-Gel 48 agarose gels longer than 30 min or E-Gel 96 agarose gels longer than 20 min.
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All E-Gel gels are labeled with expiration dates.
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Potential problems include:
- Loading too much DNA. Do not load more than the recommended amount.
- Samples with a high salt concentration. Samples containing > 50 mM NaCl, 100 mM KCl, 10 mM acetate ions, or 10 mM EDTA will cause loss of resolution.
- Samples may have been diluted in TAE instead of TE or water.
- Samples may have been pre-run; pre-running is not recommended for any of our E-Gel agarose gels.
- Sample was not properly loaded or had a very low volume of sample loaded. Load the recommended sample volume based on the gel type and loading method. For proper band separation, we recommend keeping sample volumes uniform.
- Bubbles may have been introduced while loading the samples. Bubbles will cause band distortion. Load deionized water or TE into any empty wells, and avoid introducing bubbles.
- A longer electrophoresis run time was used. Do not run the gel longer than the recommended time for each gel type. Longer run times cause an increase in the current, resulting in poor band migration or a melted gel. We recommend that you run single comb gels for 25-30 min (double comb gels 15-20 min) for straighter patterns. Do not run single comb gels longer than 40 minutes (20 minutes for double comb), or the gel will be damaged and resolution will be poor.
- Voltage or current too high. This should not be an issue with the E-Gel PowerBase power supply, which is pre-set with the proper conditions. However, researchers using the old E-Gel Base (the one that plugged into a separate power supply) should ensure that they run the gel at 60 to 70 volts (constant voltage) or 40-50 mA (constant current). Do not allow the current to exceed 60 mA.
- For E-Gel 96 Agarose Gels being run on an E-Base device, be sure you are running on the “EG” program rather than the “EP” program designed for E-PAGE gels.
- The gel was not electrophoresed immediately after sample loading (for best results, run gel within 1 min of loading).
- The gel may have been used beyond its expiration date. Check the expiration date.
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The agarose used in the 0.8%, 1.2% and 2% E-Gel agarose gels is molecular biology grade, with a normal melting point and less than 0.25% ash content. For the 4% E-Gel agarose gels, we use a special high-resolution agarose.
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The E-Gel agarose gel system uses TAE buffer.
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The dimensions of a single comb 12 well E-Gel agarose gel are as follows: The cassette itself is 8 cm by 10 cm and 0.6 cm thick. The workable gel area is 7.5 cm by 5.8 cm (from well to bottom). The thickness of the gel is estimated to be 0.4 cm. There are 12 wells and each well is 4 mm wide. The space between the wells is 1 mm.
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We do not recommend using the E-Gel Power Snap Electrophoresis Device to image pour-your-own gels.
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We do not recommend using the E-Gel Power Snap Electrophoresis Device to run pour-your-own gels.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Yes, both the E-Gel GelCapture Acquisition Software and E-Gel GelQuant Express Analysis Software applications are compatible with Windows 10; however, the E-Gel GelQuant Express Analysis Software will require the purchase of an E-Gel Imager Quantitation USB dongle (Cat. No. 4466610):
https://www.thermofisher.com/order/catalog/product/4466610
Please visit the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/e-gel-electrophoresis-system/e-gel-imager-system/e-gel-software.html) and follow the instructions to download the software.
Note: Please make sure that you download the correct version of the E-Gel GelCapture Software based on the serial number of your instrument.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Each program on the E-Gel Power Snap Electrophoresis Device is optimized to obtain the best possible results. The device does not offer user-adjustable voltage functionality.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
To select the DNA ladder that yields the best resolution for your specific E-Gel, please refer to the table on page 45 of the E-Gel Power Snap Electrophoresis System user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017050_egel_powersnapsystem_UG.pdf).
Follow recommended DNA dilutions and leave the gel to cool down on the bench or for a few minutes in the fridge. Please check troubleshooting tips provided in the manual.
No. Standard safety and hazardous waste disposal procedures should be followed when handling E-Gel EX agarose gels.
E-Gel EX agarose gels separate DNA faster, offer enhanced sensitivity, and provide added flexibility. The stain within the E-Gel EX agarose gels is SYBR Gold II which has similar spectral properties but increased sensitivity compared to SYBR Safe DNA Gel Stain. E-Gel EX agarose gels are especially suited for applications where high sensitivity is critical.
No. E-Gel agarose gels have enough buffering capacity for one run only. Performance will be impaired with multiple runs.