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View additional product information for SYBR™ Safe DNA Gel Stain - FAQs (S33102)
64 product FAQs found
很多衣服上的增白剂，以及某些真菌和细菌，和SYBR Safe DNA凝胶染料会发出同样波长的荧光。这些位于凝胶表面或凝胶内的污染物可能会产生斑点。
SYBR染料仅在很窄的pH范围内有效，约为pH 7-8。超出此范围，荧光信号将迅速消失。
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。
使用乙醇沉淀法可轻松去除核酸上的SYBR Safe DNA凝胶染料。
我们强烈不建议重复使用SYBR Safe DNA凝胶染料，因为这会大大降低灵敏度。
SYBR Safe DNA凝胶染料与目前我们已经检测的所有下游应用兼容，包括从凝胶上切割PCR产物、凝胶纯化、Gateway克隆、TOPO克隆和限制性内切酶克隆。如果您想将SYBR Safe DNA凝胶染料用于特殊应用，请将详细信息发送到我们的邮箱techsupport@lifetech.com.
SYBR Safe DNA凝胶染料有两个主要的激发峰：紫外区的280nm和可见光区的502nm。因此，254 nm或300 nm紫外激发光以及488 nm激光、470 nm LED和广泛的蓝色光源（如Safe Imager蓝光透射仪（货号S37102））都能够有效激发。最大激发波长为502nm；因此，Safe Imager蓝光透射仪是激发SYBR Safe DNA凝胶染料的最佳选择。您可通过在线方式或在染料附带的实验方案中查看SYBR Safe DNA凝胶染料的全部激发和发射光谱。
SYBR Safe DNA凝胶染料的最大发射波长为530 nm。因此一些溴化乙锭滤光片可以透过所有500 nm以上的光。这些滤光片（通常为黄色）及其相关相机设置适用于SYBR Safe DNA凝胶染料，通常只需对曝光或补光稍作调整。其他溴化乙锭滤光片（通常为红色）只能透过600 nm或以上的光；这些滤光片及其相关相机设置不适用于SYBR Safe DNA凝胶染料。
请点击此处(https://www.thermofisher.com/cn/zh/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/dna-stains/sybr-safe.html)并选择“滤光片推荐”，查看适用于SYBR Safe DNA凝胶染料的推荐滤光片。请注意，SYBR Safe DNA凝胶染料的激发和发射光谱与SYBR Green I、SYBR Green II、SYBR Gold 染料，还有荧光素（FITC）极为相似。因此，亦可使用适用于这些染料的滤光片。若使用Safe Imager蓝光透射仪，则无需使用相机滤光片；仪器内提供的琥珀色滤光片即具有该功能。
使用SYBR Safe DNA凝胶染料染色的条带在300nm透射仪上是肉眼可见的。如果相机没有配备滤镜，激发光将被捕捉到图像上，以呈现“被冲洗”的外观。在您的CCD或相机上安装紫外兼容的发射滤光片，可得到最佳的凝胶成像结果。紫外灯也会发射一些红外（IR）光；如果您的相机镜头没有可阻断IR的特殊涂层，则需要安装红外滤镜，防止凝胶后面的紫外灯形成模糊图像。为了达到最佳检测和完全安全的目的，应使用Safe Imager蓝光透射仪。
与紫外光不同，蓝光可将DNA损害降至最低。相较于使用溴化乙锭染色并暴露于紫外光的DNA，使用SYBR Safe DNA凝胶染料和Safe Imager蓝光透射仪可提高克隆效率。
不推荐。大部分深琥珀/橙色溴化乙锭滤光片在550nm附近有临界值。SYBR Green染料在520nm处有发射光，所以使用这种滤光片会过滤掉您所需要的数据。
使用SYBR Safe DNA凝胶染料染色的DNA可使用蓝光透射仪观察，如我们的Safe Imager 2.0蓝光透射仪或标准紫外透射仪。如果您想要使用DNA进行克隆，应避免使用SYBR Safe DNA凝胶染料染色的DNA暴露于紫外光照射下。
1X染料溶液中的SYBR Safe DNA凝胶染料含量低于1ppm。
可以。只需在制备融化的琼脂糖时，用SYBR Safe DNA凝胶染料溶液替代缓冲液。如果您使用了10,000X SYBR Safe DNA凝胶染料浓缩液，则使用琼脂糖凝胶缓冲液（如1X TBE或1X TAE）以1:10,000稀释浓缩染料，并将缓冲液/染料溶液加入到琼脂糖粉末中。您可使用微波炉对琼脂糖/SYBR Safe DNA凝胶染料混合物进行短暂加热。
所有SYBR染料都具有相似的光谱性质，但化学组成不同。所有SYBR染料都与dsDNA、ssDNA和RNA结合，但敏感性不同。SYBRSafe DNA凝胶染料是专门研发的溴化乙锭安全替代品。SYBR Green I是dsDNA的超灵敏染料，而SYBR Green II是RNA和ssDNA的高灵敏性染料。所有SYBR染料都适用于Safe Imager蓝光透射仪激发观察。
SYBR Safe染料无致突变性、无毒性。SYBR Safe染料与溴化乙锭在紫外范围内具有相似的敏感度（约500 pg/条带）和同等的分辨率。成像时，溴化乙锭采用标准紫外透射仪成像，而SYBR Safe染料可在紫外光（约280 nm）且具有可见光性能的激光扫描仪或蓝光（450-500 nm）下观察。
处理方法有很多种。请联系您当地的安全办公室或市政当局咨询处理指南。
所有SYBR染料都与dsDNA、ssDNA和RNA结合，但敏感性不同。SYBR Green I用于dsDNA和ssDNA染色。SYBR Green II可以对dsDNA和ssDNA进行染色，但对RNA的敏感性更好。SYBR Gold 是在SYBR Green I和II之后开发的，是最敏感的荧光凝胶染色剂。SYBR Safe DNA凝胶染色剂是一种用于蓝光系统的致突变性更低的配方。它不像SYBR Green I和II那么敏感，但可与溴化乙锭相比。 < / p >
Many whitening agents used in clothing, as well as some fungi and bacteria, fluoresce at the same wavelengths as SYBR Safe DNA gel stain. These contaminants within or on the surface of the gel may produce this speckling.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
The SYBR dyes are useful only over a narrow range of pH, from about 7 to 8. Outside this range, the fluorescent signal diminishes rapidly.
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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.
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SYBR Safe DNA Gel Stain is easily removed from nucleic acids by ethanol precipitation.
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We strongly discourage the reuse of SYBR Safe DNA Gel Stain, as this practice significantly lowers sensitivity.
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Yes. SYBR Safe DNA Gel Stain is compatible with all downstream applications we have tested so far, including excising PCR products from gels, gel purification, Gateway cloning, TOPO cloning, and restriction enzyme cloning. If you have a unique application that works with SYBR Safe DNA Gel Stain, send us the details at techsupport@thermofisher.com.
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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, wavelengths from 254 nm to 300 nm UV excitation will work, as will excitation with 488 nm lasers, 470 nm LEDs, and broad blue light sources (such as the Safe Imager Blue-Light Transilluminator (Cat. No. S37102). Maximal excitation occurs at 502 nm; the Safe Imager Blue-Light Transilluminator is therefore the best choice for excitation of SYBR Safe DNA Gel Stain. You can find the full excitation and emission spectra for SYBR Safe DNA Gel Stain online and also in the protocol provided with the stain.
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Some ethidium bromide filters allow the transmission of all light above 500 nm. These filters (which are often yellow) 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) 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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Please go here (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/dna-stains/sybr-safe.html) and click on the “Filter Recommendations” tab to see filter recommendations for use with SYBR Safe DNA Gel Stain. Note that the excitation and emission spectra of SYBR Safe DNA Gel Stain are very similar to those of SYBR Green I, SYBR Green II, and SYBR Gold dyes, as well as fluorescein (FITC). Therefore, filters appropriate for these dyes can also be used. A camera filter is not required with the Safe Imager Blue-Light Transilluminator; the amber filter provided with the instrument serves this purpose.
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Bands stained with SYBR Safe DNA Gel Stain are visible to the eye on a 300 nm transilluminator. If a camera is not equipped with a filter, the excitation light will be captured on the image to give a 'washed out' appearance. You can obtain optimum detection by photographing the gel using a UV-compatible emission filter with your CCD or film camera. UV bulbs may also emit some infrared (IR); if your camera lens is not specially coated to block IR, an IR-blocking filter is needed to prevent the appearance of faint images of the UV bulbs behind your gel. For optimum detection and complete safety, use the Safe Imager Blue-Light Transilluminator.
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Unlike UV light, blue light causes minimal damage to DNA. Use of SYBR Safe DNA Gel Stain and the Safe Imager Blue-Light Transilluminator gives improved cloning efficiency over DNA stained with ethidium bromide and exposed to UV light.
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This is not recommended. Most deep amber/orange ethidium bromide filters have a cutoff value around 550 nm. The SYBR Green dyes emit at 520 nm, which will not be detected using this filter.
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DNA stained with SYBR Safe DNA Gel Stain can be viewed using a blue-light transilluminator such as our Safe Imager 2.0 Blue-Light Transilluminator or a standard UV transilluminator. If you plan to use the DNA for cloning, avoid exposing DNA stained with SYBR Safe DNA Gel Stain to UV light.
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The SYBR Safe DNA Gel Stain content in a 1X solution is less than 1 ppm.
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Yes. Simply substitute a SYBR Safe DNA Gel Stain solution for the buffer when preparing the molten agarose. If you are using the 10,000X SYBR Safe DNA Gel Stain concentrate, dilute the concentrated stain 1:10,000 in agarose gel buffer (e.g., 1X TBE or 1X TAE) and add the buffer/stain solution to the powdered agarose. You can heat the agarose/SYBR Safe DNA Gel Stain mixture briefly in the microwave.
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All SYBR dyes have similar spectral properties, but have different chemical compositions. All SYBR dyes bind to dsDNA, ssDNA and RNA but vary in sensitivity. SYBR Safe DNA Gel Stain (Cat. No. S33102) was specifically developed as a safer alternative to ethidium bromide. SYBR Green I (Cat. No. S7585) is an ultrasensitive stain for dsDNA, and SYBR Green II (Cat. No. S7564) is a highly sensitive stain for RNA and ssDNA. All SYBR dyes are optimally excited by the Safe Imager Blue-Light Transilluminator.
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SYBR Safe Stain is non-mutagenic and non-toxic. It shows very similar sensitivity in the UV range, ~500 pg/band, with equivalent resolution to ethidium bromide. For imaging, ethidium bromide is visualized using a standard UV transilluminator, while SYBR Safe stain can be viewed with either UV (~280 nm), laser scanners with visible light capability, or blue light (450 to 500 nm).
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Disposal regulations vary. Please contact your safety office or local municipality for disposal guidelines.
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Different SYBR dyes bind to dsDNA, ssDNA, and RNA, but vary in the sensitivity and specificity with which they bind to different nucleic acids. SYBR Green I Nucleic Acid Gel Stain is used for staining dsDNA and ssDNA. SYBR Green II RNA Gel Stain will stain dsDNA and ssDNA but has better sensitivity for RNA. SYBR Gold Acid Gel Stain was developed after SYBR Green I and II and is the most sensitive fluorescent gel stain offering the highest sensitivity for both DNA and RNA. SYBR Safe DNA Gel Stain is a reduced mutagenicity formula designed for use with blue light systems. It is less sensitive than the SYBR Green I and II but comparable to ethidium bromide.
Please check the following link for additional details regarding the different SYBR dyes: The Molecular Probes Handbook: Nucleic Acid Detection on Gels, Blots and Arrays
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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 may be briefly microwaved with no loss of performance. However, we do not know the effect of repeated or very long duration microwaving.
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We strongly discourage the reuse of SYBR Safe stain, as this practice significantly lowers sensitivity.
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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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Dilute SYBR Safe stain concentrate 10,000-fold in TAE or TBE buffer prior to use. 50 mL of 1X stain is sufficient for most minigels (e.g., dilute 5 µL of concentrate into 50 mL buffer for a 1X solution). For larger gels, increase volumes proportionally, ensuring that the entire gel is fully immersed during staining.
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SYBR Safe DNA gel stain is compatible with all downstream applications we have tested so far, including excising PCR products from gels, gel purification, Gateway cloning, TOPO cloning, and restriction enzyme cloning. The use of SYBR Safe DNA gel stain with non-UV blue light emitted by the Safe Imager instrument allows you to purify DNA with virtually no UV-induced nicking or crosslinking compared to ethidium bromide and UV, resulting in dramatically increased cloning efficiencies. If you have a unique application that works with SYBR Safe DNA gel stain, send us the details at techsupport@thermofisher.com.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
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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SYBR Safe DNA gel stain was tested extensively by four independent testing laboratories. The white paper "SYBR Safe DNA Gel Stain: Assessment of mutagenicity and environmental safety" may be downloaded from our website on the 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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You can find information about all of our latest testing results on the SYBR Safe home page, which you can find by searching "SYBR Safe DNA Gel Stain" from the Thermo Fisher Scientific website home page.
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In numerous tests carried out by independent, licensed testing laboratories, SYBR Safe stain showed little or no genotoxicity and no acute toxicity. This stain is not classified as hazardous waste under U.S. federal regulations. However, please exercise common safe laboratory practice when using this reagent - wearing gloves is still recommended. For more information on safety testing of the 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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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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Yes, it is available in both buffers. The catalog numbers are as follows:
Cat. No. S33111 SYBR Safe DNA Gel Stain in 1X TAE
Cat. No. S33100 SYBR Safe DNA Gel Stain, 1L in 1X TBE
Cat. No. S33101 SYBR Safe DNA Gel Stain, 4L in 1X TBE
Cat. No. S33110 SYBR Safe DNA Gel Stain Starter Kit, includes 1L SYBR Safe DNA Gel Stain (Cat. No. S33100) in 1X TBE and one photographic filter (Cat. No. S37100)
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Yes, the agarose can be prepared directly in the SYBR Safe DNA gel stain (it may be heated in microwave). Run the gel normally, no destaining is required.
*Note: SYBR Safe DNA gel stain is prepared 0.5X TBE buffer. Use this preparation directly when making up a gel. Gels that include SYBR Safe DNA gel stain can be run in 0.5X or 1X TBE buffer without effects on resolution.
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SYBR Safe DNA gel stain was developed specifically for reduced mutagenicity to be safer than ethidium bromide for staining DNA in agarose or acrylamide gels. SYBR Safe DNA gel stain is not only less mutagenic than ethidium bromide, but its detection sensitivity is better than that of ethidium bromide.
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SYBR Safe DNA Gel Stain (Cat. No. S33102) can be used in place of ethidium bromide for all staining applications, including RNA staining.
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Water and 70% ethanol should get rid of most stains from spills. For persistent stains, bleach may be used. Use a handheld UV light in the darkroom to check for any remaining stain.
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Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.
Yes, the Dual LED Blue/White Light Transilluminator can be used to visualize gels stained with SYBR Safe DNA Gel Stain, SYBR Gold Nucleic Acid Gel Stain, or Ethidium bromide.
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Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.