Denaturing urea polyacrylamide gel electrophoresis (Urea PAGE)
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Updated: Jun 19, 2026

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)
Published on: October 29, 2009
Heike Summer1, René Grämer, Peter Dröge
1School of Biological Sciences, Nanyang Technological University, Singapore - NTU. summer@ntu.edu.sg
Abstract:
Urea PAGE or denaturing urea polyacrylamide gel electrophoresis employs 6-8 M urea, which denatures secondary DNA or RNA structures and is used for their separation in a polyacrylamide gel matrix based on the molecular weight. Fragments between 2 to 500 bases, with length differences as small as a single nucleotide, can be separated using this method(1). The migration of the sample is dependent on the chosen acrylamide concentration. A higher percentage of polyacrylamide resolves lower molecular weight fragments. The combination of urea and temperatures of 45-55 degrees C during the gel run allows for the separation of unstructured DNA or RNA molecules. In general this method is required to analyze or purify single stranded DNA or RNA fragments, such as synthesized or labeled oligonucleotides or products from enzymatic cleavage reactions. In this video article we show how to prepare and run the denaturing urea polyacrylamide gels. Technical tips are included, in addition to the original protocol (1,2).
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