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Estimation of binding constants of peptide nucleic acid and secondary-structured DNA by affinity capillary electrophoresis.
- Source :
-
Analytical chemistry [Anal Chem] 2012 Jun 19; Vol. 84 (12), pp. 5204-9. Date of Electronic Publication: 2012 Jun 05. - Publication Year :
- 2012
-
Abstract
- An affinity capillary electrophoresis method was developed to determine a binding constant between a peptide nucleic acid (PNA) and a hairpin-structured DNA. A diblock copolymer composed of PNA and polyethylene glycol (PEG) was synthesized as a novel affinity probe. The base sequence of the probe's PNA segment was complementary to a hairpin-structured region of a 60-base single-stranded DNA (ssDNA). Upon applying a voltage, the DNA hairpin migrated slowly compared to a random sequence ssDNA in the presence of the PNA probe. This retardation was induced by strand invasion of the PNA into the DNA hairpin to form a hybridized complex, where the PEG segment received a large amount of hydrodynamic friction during electrophoresis. The binding constant between the PNA probe and the DNA hairpin was easily determined by mobility analysis. This simple method would be potentially beneficial in studying binding behaviors of various artificial nucleotides to natural DNA or RNA.
- Subjects :
- Affinity Labels chemical synthesis
Affinity Labels chemistry
Affinity Labels metabolism
Base Sequence
DNA genetics
Inverted Repeat Sequences
Nucleic Acid Hybridization
Polyethylene Glycols chemistry
Thermodynamics
DNA chemistry
DNA metabolism
Electrophoresis, Capillary methods
Nucleic Acid Conformation
Peptide Nucleic Acids metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1520-6882
- Volume :
- 84
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Analytical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 22624952
- Full Text :
- https://doi.org/10.1021/ac301025m