Deprotection of oligonucleotides and purification using denaturing PAGE

Curr Protoc Immunol. 2001 May;Chapter 10:Unit 10.7. doi: 10.1002/0471142735.im1007s06.

Abstract

The advantages of purification by denaturing polyacrylamide gel electrophoresis (PAGE) are speed, simplicity, and high resolution. Although yields tend to be low (<50% of applied sample), the amount of material recovered is usually far in excess of that required for most molecular biology applications (e.g., cloning and sequencing). Denaturing PAGE can resolve oligonucleotides from 2 to 300 bases, depending on the percentage of polyacrylamide used. The method presented here is thus useful not only for isolating chemically synthesized deoxyribonucleotides but also small RNAs or other single-stranded polynucleotides.

Publication types

  • Review

MeSH terms

  • Cloning, Molecular / methods
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Oligonucleotides / chemical synthesis
  • Oligonucleotides / chemistry
  • Oligonucleotides / isolation & purification*
  • Sequence Analysis, DNA / methods

Substances

  • Oligonucleotides