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should i add EtBr in the gel solution, when i run the mRNA?
thanks!
thanks!
lol, that's funnyMoonbear said:then again, I'm not yet old enough to be one of those profs the students chase out of the lab...I've worked for those, the ones who make a big mess when they come into the lab, so everyone chases them out as quickly as possible if they get the urge to do some benchwork.
Monique said:it's not much of an effort to melt 1g of agarose in 100 ml of buffer
EtBr (Ethidium Bromide) is a fluorescent dye commonly used in molecular biology for visualizing nucleic acids, such as mRNA, in gel electrophoresis. It intercalates between the base pairs of DNA or RNA, causing it to fluoresce under UV light. This allows for the visualization and quantification of nucleic acids in the gel.
EtBr does not significantly affect the migration of mRNA in the gel during electrophoresis. Instead, it is used to visualize the mRNA after it has been separated based on size. The migration of the mRNA is primarily determined by the size and charge of the molecule, as well as the strength of the electric field.
EtBr is considered to be a hazardous chemical and should be handled with caution. It is a mutagen and a potential carcinogen, so it is important to wear appropriate personal protective equipment (PPE) when handling it. Additionally, proper disposal methods should be followed to prevent environmental contamination.
Yes, there are alternative dyes that can be used for visualizing mRNA in gels, such as SYBR Green or GelRed. These dyes have similar mechanisms of action to EtBr and are considered to be safer alternatives. However, they may have different sensitivity or specificity compared to EtBr, so it is important to optimize their use for each experiment.
One limitation of using EtBr in gel electrophoresis is that it can only detect the presence of nucleic acids, but not their sequence or specific mutations. Additionally, the sensitivity of EtBr may vary, making it difficult to accurately quantify the amount of mRNA in a sample. It is also important to note that EtBr may interfere with downstream applications, such as cloning or sequencing, so alternative methods of mRNA analysis may be necessary in these cases.