I am setting up my lab at a new institution and I am being encouraged to use GelRed for Agarose Electophoresis due to its safety. I have been told that, although the dye works, it usually alters the molecular weight of the bands and this is unacceptable for cloning experiments.
I am wondering if anyone here has experience with this dye and what has been their experience.
On other hand, many years ago I was taught that the Cancer risk associated with Ethidium Bromide comes from its property as an intercalating agent. GelRed is also an intercalating agent, therefore, it is logical to deduce that they might be equally risky. The manufacturers say that they are safe based on results of the Ames test. Is there any additional and independent evidence probing its safety?
Hello, I’m a scientist at Biotium and would like to provide you with some information about GelRed that answers both of these commonly asked questions:
1. "I have been told that, although the dye works, it usually alters the molecular weight of the bands and this is unacceptable for cloning experiments."
2. "Many years ago I was taught that the Cancer risk associated with Ethidium Bromide comes from its property as an intercalating agent. GelRed is also an intercalating agent, therefore, it is logical to deduce that they might be equally risky"
GelRed was designed with safety in mind, and therefore is a larger molecule than EtBr. This size means that is unable to cross healthy cell membranes and therefore unable to reach the DNA in living cells. It’s safety has been independently tested and verified and that information can be found here:
http://biotium.com/wp-content/uploads/2013/07/GR-GG-Safety.pdf
The fact that it is a larger dye also speaks to your first question. Because of its larger size, it can impact migration of DNA fragments when used as a pre-cast stain. Therefore, when very accurate information is needed about the size of a band, we recommend that researchers use the post-staining method, which can be performed successfully in as little as 10 minutes.
As with EtBr, GelRed can be used for cloning and is removed from DNA during the purification process with a standard gel extraction kit.
Here is some more food for thought: using a blue-light excitation source with a green dye like GelGreen gives as much as 100 to 1000-fold improvements in cloning efficiency compared to using a UV transilluminator by preventing UV damage to DNA during band excision from the gel.
https://www.researchgate.net/publication/14283779_Protection_of_DNA_during_preparative_agarose_gel_electrophoresis_against_damage_induced_by_ultraviolet_light?ev=prf_pub
I'll be happy to answer any other questions or concerns you may have about using GelRed or our other products!
http://biotium.com/wp-content/uploads/2013/07/GR-GG-Safety.pdf
Article Protection of DNA During Preparative Agarose Gel Electrophor...
I have been using it and its working fine. Not sure about it being less toxic and its also an i intercalating nucleic acid stain like ethidium bromide. The substance is marketed as a less toxic and more sensitive alternative to ethidium bromide. Good luck
GelRed has low toxicity because it can't pass through cell membranes, but still has some toxicity... in our lab we have the same safety protocol for GelRed zone that we had with EtBr, just in case. We have not detected any problem with molecular weight...
Thanks for your answers! I really appreciate your help! I´ll have your considerations in mind.
Hello, I’m a scientist at Biotium and would like to provide you with some information about GelRed that answers both of these commonly asked questions:
1. "I have been told that, although the dye works, it usually alters the molecular weight of the bands and this is unacceptable for cloning experiments."
2. "Many years ago I was taught that the Cancer risk associated with Ethidium Bromide comes from its property as an intercalating agent. GelRed is also an intercalating agent, therefore, it is logical to deduce that they might be equally risky"
GelRed was designed with safety in mind, and therefore is a larger molecule than EtBr. This size means that is unable to cross healthy cell membranes and therefore unable to reach the DNA in living cells. It’s safety has been independently tested and verified and that information can be found here:
http://biotium.com/wp-content/uploads/2013/07/GR-GG-Safety.pdf
The fact that it is a larger dye also speaks to your first question. Because of its larger size, it can impact migration of DNA fragments when used as a pre-cast stain. Therefore, when very accurate information is needed about the size of a band, we recommend that researchers use the post-staining method, which can be performed successfully in as little as 10 minutes.
As with EtBr, GelRed can be used for cloning and is removed from DNA during the purification process with a standard gel extraction kit.
Here is some more food for thought: using a blue-light excitation source with a green dye like GelGreen gives as much as 100 to 1000-fold improvements in cloning efficiency compared to using a UV transilluminator by preventing UV damage to DNA during band excision from the gel.
https://www.researchgate.net/publication/14283779_Protection_of_DNA_during_preparative_agarose_gel_electrophoresis_against_damage_induced_by_ultraviolet_light?ev=prf_pub
I'll be happy to answer any other questions or concerns you may have about using GelRed or our other products!
http://biotium.com/wp-content/uploads/2013/07/GR-GG-Safety.pdf
Article Protection of DNA During Preparative Agarose Gel Electrophor...
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