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View additional product information for E-Gel™ Agarose Gels with SYBR™ Safe DNA Gel Stain - FAQs (A45202, A45203, A45204, A45205, A45206, A42136, A42135, A42100)
42 product FAQs found
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。
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。
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。
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within ourNucleic Acid Gel Electrophoresis and Blotting Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Gel Electrophoresis and Blotting Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Gel Electrophoresis and Blotting Support Center.
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.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Pre-run is no longer necessary for any of our E-Gel agarose gels.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
All E-Gel gels are labeled with expiration dates.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within ourNucleic Acid Gel Electrophoresis and Blotting Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
The E-Gel agarose gel system uses TAE buffer.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
We do not recommend using the E-Gel Power Snap Electrophoresis Device to image pour-your-own gels.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Due to the differences in dimensions, the new E-Gel 11-well and 22-well agarose gel formats are not compatible with the E-Gel Simple Runner Electrophoresis Device.
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.