Separation of DNA Oligonucleotides Using Denaturing Urea PAGE

Denaturing urea polyacrylamide gel electrophoresis (PAGE) allows the separation of linear single-stranded DNA molecules based on molecular weight. This method can be used to analyze or purify short synthesized DNA oligonucleotides or products from enzymatic reactions.In this chapter we describe how to prepare and how to run these high concentration polyacrylamide gels. We detail how to transfer a gel onto Whatman paper and how to dry it. Radiolabelled oligonucleotides are visualized by PhosphorImager technology.