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Specifications
Print Version
| Description | SafeView |
|---|---|
| SKU | G468 |
| SafeView Series | Post Staining Dye |
| LED Viewer Compatibility | Yes |
| Stain Color | Green |
| Applications | Safe Detection of dsDNA, ssDNA and RNA in agarose and polyacrylamide gels. |
| Note | All SafeView DNA Stains are ISO-13485 certified. Dispose of SafeView DNA Stains as you would any other non-carcinogenic fluorescent dye (eg. Acridine orange; Propidium iodide). |
| Shipping Conditions | Shipped on blue ice packs. |
| Storage Condition | Store at 4°C for up to 2 years. |
| Unit quantity | 1.0 ml (10,000X) |
Documents
- SafeView Plus Protocol
- SafeView Plus™
- Sensitivity Test
- SafeView Plus Promotional Flyer
FAQs
| Can SafeView be a replacement for ethidium bromide? Can I do gel extraction with it? | |
SafeView can be used as a replacement for ethidium bromide as they both work on general agarose. We recommend using SafeGreen for downstream cloning applications as SafeView can interfere with the ligation reaction, yielding fewer colonies. | |
| How does SafeView work and why is it not carcinogenic? | |
There are fluorescent compounds in SafeView and these fluorescent compounds have the capability to bind to DNA. There may be some unknown effects of SafeView that have not been documented but that applies to the SYBR set as well; however, SafeView products are definitely not as carcinogenic as ethidium bromide. | |
| How do I use SafeView products? | |
The Safe-(Red, Green and White) loading dyes work the same as 6x loading dye, loaded with the sample. SafeView Classic is used directly in the gel and the running buffer. | |
| Does the SafeView differentiate double stranded nucleic acid and single stranded nucleic acid? Does the Safe-(Red, Green and White) work the same way? | |
Under UV, SafeView Classic emits a green fluorescence when bound to both single and double stranded DNA templates, therefore they cannot be differentiated by this method. It will emit a red fluorescence when bound to RNA templates.The SafeView Stains (Red, Green and White) do not perform in this way and will stain all nucleic acid templates one color. | |
| At what temperature do I store the SafeView products? | |
All the SafeView products should be stored at four degrees Celsius. | |
| Do I need a special filter for photography of DNA gels stained with SafeView? | |
Under UV light, SafeView Classic emits a green fluroescence when bound to both single and double stranded DNA templates. It will emit a red fluorescence when bound to RNA templates. No filter is necessary for viewing these colours, however a filter may be needed for photographing the gel. | |
| How long does the SafeView Classic stain last in a gel? | |
Our in-house testing has shown that SafeView stained gels (>10ng DNA loaded per lane) can still be effectively visualized up to 1 week later with only a slight decrease in brightness. Gels should be stored properly to maintain a good signal, at 4C in the dark, sealed in a plastic bag or pouch with wet paper towel loosely wrapped around the gel. | |
| Why is SafeView (G-108) stain not working on my samples? | |
Make sure you are following the protocol carefully. It is critical that both buffer and gel have SafeView dye in them otherwise it will not work. This is different from ethidium bromide. You can consider to add 2.5ul of SafeView (instead of 5ul) for every 100ml of running buffer, which will reduce background fluorescence and allow the bands to show with more contrast. | |
| Is it degradable, if so how fast and under what circumstances? | |
2 hours over 100C | |
| What is the sensitivity of the dyes? | |
Safe-Green has Excitation Wavelength of 490nm and Emission Wavelength of 525nm, and its sensitivity range is between 0.2-0.6ng.Safe-Red has Excitation Wavelength of 540nm and Emission Wavelength of 630nm, and its sensitivity range is between 0.3-0.8ng.Safe-White has Excitation Wavelength of 370nm and Emission Wavelength of 470nm, and its sensitivity range is between 0.2-0.5ng.SafeView Classic emits green fluorescence when bound to dsDNA and ssDNA and red fluorescence when bound to RNA. This stain has one excitation (490 nm) and two emission spectra (520 nm and 635 nm) and the sensitivity range is between 0.1-0.3ng.SafeView Plus has Excitation Wavelength of 490nm and Emission Wavelength of 525nm and its sensitivity range is between 0.05-0.1ng. | |
| Can SafeView products be used post-stain? | |
Only SafeView Plus (G468) should be used in a post stain. SafeView classic and Safe Stains are not designed for post-staining. SafeView Classic must be added to the gel and the running buffer prior to the loading of the samples. Safe-(Red, Green and White) stains must be added to the sample before loading it to the gel. | |
| Why is the EtBr signal stronger in the pictures when I compare SafeView with EtBr? | |
A reason for this is that most gel doc systems have been optimized for EtBr so that is why the EtBr signal may be stronger in the pictures. | |
| will I need an additional loading buffer for my samples? | |
The loading dye is included in the products. No additional loading buffer needed. | |
| We see migrations and band shifting of our fragments. Are there any recommendations that you can give us to minimize this band shifting? | |
Shifting is unavoidable and quite natural for any fragments, regardless of the staining agent. We suggest to use SafeGreen ladders, which will give accurate molecular weight with no additional staining agents needed.http://www.abmgood.com/DNA-Ladder-Safe-Green™-100bp-Opti-DNA-Marker-Invitroge-G473.htmlhttp://www.abmgood.com/DNA-Ladder-Safe-Green™-1kb-Opti-DNA-Marker-Invitroge-G474.html | |
| We see shifting and migration of the DNA fragments. What are the recommendations to minimize this? | |
Shifting is unavoidable and quite natural for such fragments, regardless of the staining. We suggest using SafeGreen ladders, which will give accurate molecular weight with no additional staining agents needed.http://www.abmgood.com/DNA-Ladder-Safe-Green™-100bp-Opti-DNA-Marker-Invitroge-G473.htmlhttp://www.abmgood.com/DNA-Ladder-Safe-Green™-1kb-Opti-DNA-Marker-Invitroge-G474.html | |
| I cannot see 100bp and 200bp bands on a 1% gel. What should I do? | |
It is very difficult to detect 100bp and 200bp bands in 1% gel with any stains. Higher gel concentrations should be used, such as 2% agarose. | |
| Can I use SafeView products in Polyacrylamide gels? | |
Yes, we have tested our SafeView products for this application. | |
| Which of the Safe stains will work with blue light / LED? | |
All of our SafeView stains have been tested in-house to be compatible with UV light. SafeView Classic, SafeView Plus, and SafeGreen will also work under blue light/LED. SafeRed and SafeWhite will only work under UV light.Safe View Plus should only appear green. SafeView Classic will be red for RNA and green for DNA. | |
| What is the approved filter for eliminating the spent running buffer solutions? | |
Most facilities have approved the disposal of SafeView chemicals directly down the drain, with adequate water flushing or dilution, since SafeView™ products are non-carcinogenic. However, regulations vary between different regions, so please contact your local safety office for disposal guidelines specific to your area. You may review the MSDS of this product for more detailed information. | |
References
- Dlusskaya, E. A., Atrazhev, A. M., & Ashbolt, N. J. "Colloid chemistry pitfall for flow cytometric enumeration of viruses in water" Water Research X 2:100025 (2019). DOI: 10.1016/j.wroa.2019.100025.
- Pereira, B. A., Zangeronimo, M. G., Castillo-Martín, M., Gadani, B., Chaves, B. R., Rodríguez-Gil, J. E., … Yeste, M. "Supplementing Maturation Medium With Insulin Growth Factor I and Vitrification-Warming Solutions With Reduced Glutathione Enhances Survival Rates and Development Ability of in vitro Matured Vitrified-Warmed Pig Oocytes" Frontiers in Physiology 9: (2019). DOI: 10.3389/fphys.2018.01894.
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放入前应加好反应体系各组分,再放入 PCR 离心管。
3、反向重复第二步骤将顶盖板向后拉,盖好顶盖。
4、按 F2“Create”新建一个扩增的 PCR 程序。
5、按控制台面右方上下左右方向键,可随意移动编辑位置,输入需要
的温度、时间、循环数等。
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7、按 F1“Start”启动
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3、反向重复第二步骤将顶盖板向后拉,盖好顶盖。
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7、按 F1“Start”启动
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9、扩增完成后按台面左方“Stop”键退出。
10、按“Hist”,可查看上次扩增的历史记录。11、完全退出后,按台面左下方电源开关,拔出插销,切断电源。
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pcr仪使用时,在设定反应程序时,一共分了多少个步骤 123
未00102021-08-05
1.常规程序将PCR反应所需的成分配置完后,在PCR仪上于94-96℃预加热几十秒至几分钟,使模板DNA充分变性,然后进入扩增循环。在每一个循环中,先于94℃保持30秒钟使模板变性,然后将温度降到复性温度(一般50-60℃之间),一般保持30秒钟,使引物与模板充分退火;在72℃保持1分钟(扩增1kb片段),使引物在模板上延伸,合成DNA,完成一个循环。重复这样的循环25~35次,使扩增的DNA片段大量累积。最后,在72℃保持3-7min,使产物延伸完整,4℃保存。2.复性(退火)和延伸温度复性的温度是PCR扩增是否顺利的关键因素,通常在50-60℃之间。具体的温度主要由引物的Tm值决定。延伸温度绝大多数设定为72℃。如果复性的温度很高,可以将延伸温度和复性温度设置成同一温度,变成二步法PCR。3.反应时间变性步骤一般使用30秒钟,如果模板的G+C含量较高,或直接用细胞做模板,变性时间可适当延长。复性时间有30秒种一般是足够的。延伸时间由扩增产物的大小决定,一般采用1kb用1分钟来保证充足的时间。4.循环次数循环次数主要与模板的起始数量有关,在模板拷贝数为104~105数量级时,循环数通常为25~35次。平台效应(plateaueffect):PCR扩增过程后期会出现的产物的积累按减弱的指数速率增长的现象。原因:底物和引物的浓度已经降低,dNTP和DNA聚合酶的稳定性或活性降低,产生的焦磷酸会出现末端产物抑制作用,非特异性产物或引物的二聚体出现非特异性竞争作用,扩增产物自身复性,高浓度扩增产物变性不彻底。5.PCR反应液的配制PCR反应体系的配置方式有时也会影响反应的正常进行。常规方法与其它酶学反应一样,在最后加入DNA聚合酶。早期的PCR仪没有带加热的盖子,要求在反应液上覆盖一层矿物油,防止水分蒸发。对于使用具3’-5’外切活性的高温DNA聚合酶时,有时会扩增不出产物。在遇到这个问题时,如果将反应成分分开配制,A管含模板、引物和dNTP,以及调整体积的H2O,B管含缓冲液、DNA聚合酶和水,然后再将两管溶液混合起来,可较好地克服这个问题。按照常规的方法配制反应体系,有时会出现非特异性扩增的问题。热启动(hotstart)PCR操作方式可较好解决这一问题。将dNTP、缓冲液,Mg2+和primer先配制好,然后加入一粒蜡珠(如AmpliWaxPCRQam100),加热熔化,再冷却,使蜡将溶液封住,最后加入模板和DNA聚合酶等剩余成分。只有当PCR反应进入高温阶段后,蜡层熔化,所有反应成分才会混合在一起。
详述PCR仪的操作使用说明和注意事项123
2021-07-25
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【马干细胞因子(SCF)ELISA试剂盒】 太平洋安防网123
m8848132021-07-21
进口的PCR仪是不是都要五六七万,或许还不止?
国产的我查了博日的,也要三五万,最小的也要3万。这是不是pcr的普遍报价阿,觉得好贵呀。
国产的质量如何?还有其他可靠的厂家吗?国产的对实验影响会有多大呀?
国产的我查了博日的,也要三五万,最小的也要3万。这是不是pcr的普遍报价阿,觉得好贵呀。
国产的质量如何?还有其他可靠的厂家吗?国产的对实验影响会有多大呀?

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