Premium
Analysis and purification of peptide nucleic acids by denaturing PAGE
Author(s) -
Dodd David W.,
Hudson Robert H. E.
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700192
Subject(s) - peptide nucleic acid , oligomer , peptide , nucleic acid , chromatography , chemistry , polyacrylamide gel electrophoresis , polyacrylamide , coomassie brilliant blue , electrophoresis , silver stain , gel electrophoresis , staining , biochemistry , microbiology and biotechnology , biology , organic chemistry , polymer chemistry , genetics , enzyme
A flexible and convenient protocol for the analysis and purification of peptide nucleic acid (PNA) oligomers and PNA‐peptide chimeras by denaturing PAGE is described. Vertical slab gel electrophoresis, 26% in polyacrylamide and 8 M urea at pH 3, was suitable for analysis of oligomers ranging in size from tetramers (4‐mers) to tetradodecamers (24‐mers). Single‐base resolution of oligomers was achieved and separations are generally superior to those given by standard RP‐HPLC techniques. The separation of a related series of PNA oligomers showed the distance migrated was linearly dependent on the logarithm of the molecular weight. The migration of oligomers through the gel is dependent on the number of basic functional groups present, such as amino groups, and the A and C content of the oligomer. PNAs are amenable to detection by UV‐shadowing technique illuminated at 260 nm or Coomassie blue staining, both with similar, sub‐microgram per band detection limits.