Ethidium Bromide Solution (0.625 mg/mL) - FAQs

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4 个常见问题解答

你们是否有在琼脂糖凝胶中进行溴化乙锭染色的实验方案？

用于琼脂糖凝胶：
•将溴化乙锭加到融化的琼脂糖中，至终浓度为0.5 µg/mL。不要融化已经含有溴化乙锭的琼脂糖。

用于电泳后的琼脂糖凝胶染色：
•在不含溴化乙锭的情况下进行电泳后凝胶染色时，可将凝胶浸泡在含0.5 µg/mL溴化乙锭的水溶液中，并轻轻搅拌10-30分钟。
•如有需要，可将凝胶置于水中摇动30分钟以脱色。对RNA凝胶染色时，应尽量缩短染色时间，并一定要进行脱色处理。

溴化乙锭可以用来检测哪些靶标？灵敏性如何？

溴化乙锭可用于检测ssDNA、RNA和dsDNA。这种荧光染料可插入到堆叠的核酸碱基间，在590 nm处产生较强的荧光。在1块琼脂糖凝胶中，溴化乙锭可检测低至约1–10 ng/条带的dsDNA。

Do you have a protocol for ethidium bromide staining in agarose gels?

For use in agarose gels:

- Add ethidium bromide to melted agarose to a final concentration of 0.5 µg/mL. Do not melt agarose that already contains ethidium bromide.

For staining agarose gels after electrophoresis:

- You can stain gels that have been run in the absence of ethidium bromide by covering the gel in 0.5 µg/mL ethidium bromide in water and gently agitating for 10 to 30 minutes.
- If necessary, gels can be destained by shaking in H2O for an additional 30 minutes. To stain RNA gels, you should minimize staining time and definitely include a destaining period.

What can ethidium bromide be used to detect and what is its sensitivity?

Ethidium bromide can be used to detect ssDNA, RNA, and dsDNA. This fluorescent dye intercalates between the stacked bases of nucleic acids, and exhibits an increased fluorescence at 590 nm. Ethidium bromide can detect down to ~1-10 ng/band of dsDNA in an agarose gel.