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Real-time enzyme-digesting identification of double-strand DNA in a resonance-cantilever embedded micro-chamber.
- Source :
- Lab on a Chip; 2014, Vol. 14 Issue 6, p1206-1214, 9p
- Publication Year :
- 2014
-
Abstract
- A novel direct identification of double-strand DNA is proposed by using real-time enzyme-digestion in a resonant-cantilever embedded microfluidic chip. The new gene-level detection method is expected to replace the conventional DNA-hybridization based gene-detection that suffers from not only nonspecific adsorption induced false-positives but also complicated single-strand DNA preparation and hybridization. Since a detected DNA chain features a unique cutting site for a certain restriction-enzyme, the accurately cut-off mass (representing the length of the digested segment) can be online recorded by the frequency-shift signal of the resonant micro-cantilever sensor. This enzyme-digestion technique is confirmed by experimental identification of the stx2 gene of E. coli O157:H7. The direct-PCR sample is directly analyzed by using our lab-made cantilever-embedded microfluidic-chip. The 3776 bp DNA is immobilized via biotin–streptavidin binding and the added mass is recorded by a frequency-decrease of 15.9 kHz within 10 min. Then, with EcoRV-enzyme digestion at the site of 2635 bp, the cut-off mass is real-time detected by a frequency-increase of 10.2 kHz within 6 min. The detected frequency-shift ratio of 15.9/10.2 = 64.2% is consistent with the length ratio between the cut-off fragment and the whole DNA chain (2635/3776 = 69.8%). Hence, the simple and accurate double-strand detection method is verified experimentally. [ABSTRACT FROM AUTHOR]
- Subjects :
- DNA
RESONANCE
CANTILEVERS
STREPTAVIDIN
SPECTRUM allocation
Subjects
Details
- Language :
- English
- ISSN :
- 14730197
- Volume :
- 14
- Issue :
- 6
- Database :
- Complementary Index
- Journal :
- Lab on a Chip
- Publication Type :
- Academic Journal
- Accession number :
- 100403972
- Full Text :
- https://doi.org/10.1039/c3lc51294k