I have a question because recently my lab changed Ethidium Bromide to GelRed Nucleic acid stain, from Biotium cat: 41003F. They said that is the same and also less dangerous. But people from another lab next to me said that is not that sensitive and you need big amount of DNA to get good results.. my case is that the organism that I work with has little amount of everything and now I'm having some problems with some PCR, it seems that I don't get amplification so I don't know what is the problem, maybe is a new substance. If someone has experience with this kind of things, please tell me to get an idea.
Hi, Maria, GelRed have even more sensitivity to compare with Ethidium Bromide. I think it is a problem with PCR. I suggest you to use some kind of positive control for PCR and for staining. For example, by loading several dilution of the molecular weight marker, This simple give your answer about the problem. Good luck!
Hi, I use gel red in lab and i've not problem.
1. Try to use, with the same PCR products, an electrophoresis gel with Etidium and one with GelRed in order to compare sensitivity. Eg. 10ul of product in Etidium and 10ul of the same product in GelRed.
A question...before change Etidium-GelRed..have you ever some problem with revelation in electrophoresis?
2. You can try to load in electrophoresis (with GelRed), a sample that you are sure to have an amplificated. If you reveal this product in gel with GelRed, is not a GelRed problem!
3. Try to increase GelRed amount in your Agarose Gel or varying dilution of stock GelRed or its Work solution.
I hope to help you
Mario
Hi, Maria, GelRed have even more sensitivity to compare with Ethidium Bromide. I think it is a problem with PCR. I suggest you to use some kind of positive control for PCR and for staining. For example, by loading several dilution of the molecular weight marker, This simple give your answer about the problem. Good luck!
Hi Maria, I agree with Taras, that your problem is more likely to be due to your PCR. Have a look at this article. They have done pretty much what Mario suggests (it's from a company rep, but looks legit and very relevant to your question): http://www.labnews.co.uk/features/a-toxic-death-for-ethidium-bromide/
Good luck!
Thank you guys!! It was really helpful ur answer!! im going to do all the advices that i got... thank you!!! and I wish you good luck also!!!
Have you change the light source? Although EtBr uses UV light for being observed, GelRed need blue light…, slightly different wavelength but this may contribute to the less sensitivity that some people may observe after changing to the new DNA staining agent.
- Badges
- Science method