RNA Markers
RNA Loading Dye That Enables RNA Electrophoresis on Non-denaturing Agarose Gel
RNA Ezloading Dye
Highlights
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Just mix with RNA and heating.
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RNA can be separated by electrophoresis using formaldehyde-free agarose gel with TAE/TBE buffer.
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RNA is stained with Ethidium bromide (EtBr) during denaturation step. No need for gel staining.
Product Information
Single-stranded RNA molecules form hydrogen bonds between their bases and adopt higher-order structures. This makes it difficult to determine the RNA mobility reflecting molecular size in agarose gel electrophoresis. Therefore, denaturing agarose gel electrophoresis is generally performed to suppress hydrogen bonding within RNA molecules. In this method, denaturing agent is added to the agarose solution, and formaldehyde is commonly used as a denaturant. Formaldehyde is toxic and requires careful handling.
Here, RNA Ezloading Dye enables RNA electrophoresis using formaldehyde-free agarose gels. This innovative reagent can eliminate the time spent preparing denaturing agarose gels and simultaneously reduce the psychological burden on experimenters caused by using large amounts of formaldehyde solution.
Procedure Overview
(1) Mix RNA Ezloading Dye and 37% formaldehyde solution in a 19:1 ratio.
Ex. RNA Ezloading Dye 19 μL ＋ Formaldehyde solution 1 μL
(2) Mix (1) and RNA sample in a 1:1 ratio.
Ex. Solution (1) 2.5 μL ＋ RNA sample 2.5 μL
(3) Incubate (3) at 75℃ for 3 minutes for RNA denaturation. In this step, RNA is stained with EtBr.
(4) Load the sample onto a formaldehyde-free agarose gel.
Application
Fig.1 RNA marker (#DM160) or RNA sample (Human Total RNA) was electrophoresed using RNA Ezloading Dye. RNA samples prepared using RNA Ezloading Dye can be separated by electrophoresis with a formaldehyde-free agarose gel. And the mobility of each RNA on denaturing gel or non-denaturing gel didn’t change.
Fig.2 RNA markers (#DM160) was electrophoresed using RNA Ezloading Dye or similar products of other companies according to each protocols with a formaldehyde-free agarose gel and TAE buffer. Using RNA Ezloading Dye, sharp RNA bands were obtained without ethidium bromide staining.
The mobility of RNA prepared using RNA Ezloading Dye was unchanged when run on a formaldehyde-free agarose gel versus a formaldehyde-containing agarose gel (Fig.1).
Note
Formaldehyde solution and a block incubator are required for this experiment.
This product is prepared for electrophoresis using non-denaturing agarose gel, but for rigorous experiment, please also consider using denaturing agarose gel electrophoresis.
EN
Related Product：RNA High for Easy Electrophoresis
Set of RNA Ezloading Dye(#DM172) and RNA Marker(#DM160).
BDL is a member of the Funakoshi Group.
