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A mobility shift assay for DNA detection using nanochannel gradient electrophoresis
Author(s) -
Startsev Michael A.,
Ostrowski Martin,
Goldys Ewa M.,
Inglis David W.
Publication year - 2017
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.201600358
Subject(s) - peptide nucleic acid , oligonucleotide , dna , nucleic acid , biomolecule , chemistry , electrophoresis , nucleic acid thermodynamics , nanofluidics , multiple displacement amplification , microbiology and biotechnology , chromatography , nanotechnology , biochemistry , polymerase chain reaction , biology , gene , base sequence , materials science , dna extraction
Conventional detection of pathogenic or other biological contamination relies on amplification of DNA using sequence‐specific primers. Recent work in nanofluidics has shown very high concentration enhancement of biomolecules with some degree of simultaneous separation. This work demonstrates the combination of these two approaches by selectively concentrating a mobility‐shifted hybridization product, potentially enabling rapid detection of rare DNA fragments such as highly specific 16S ribosomal DNA. We have performed conductivity gradient electrofocusing within nanofluidic channels and have shown concentration of hybridized peptide nucleic acids and DNA oligomers. We also show selectivity to single base‐pair mismatch on 18‐mer oligos. This approach may enable sensitive optical detection of small amounts of DNA.