Isothermal sensitive detection of microRNA using an autonomous DNA machine recycling output as input

Bioorganic and Medicinal Chemistry Letters

Volumn 20, Issue 20, 2010, Pages 6056-6060

  Ogawa, Atsushi ^a  

^a EHIME UNIVERSITY   (Japan)

Author keywords

Biosensor; DNA machine; MicroRNA; Molecular beacon; Nucleic acid detection

Indexed keywords

DNA; MICRORNA;

ARTICLE; GENE AMPLIFICATION; ISOTHERM; SENSITIVITY AND SPECIFICITY;

ANIMALS; BASE SEQUENCE; BIOSENSING TECHNIQUES; DNA; LIMIT OF DETECTION; MICE; MICRORNAS; NUCLEIC ACID AMPLIFICATION TECHNIQUES; TEMPERATURE;

EID: 77956916937     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.08.055     Document Type: Article

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18. This band is much thicker than a band corresponding to DM-template, indicating that non-nicked products were produced much more than DM-template. This is probably because the output DNA somewhat replaced the saturated non-nicked products on DM-template and extended by Bst DNA polymerase to be non-nicked products. Although the overproduced single-stranded non-nicked products have potential to open the MB-probe as well as the output DNA, the amount is estimated to be much smaller (∼10%) compared to that of the output DNA even in the presence of 500 amol of target RNA. Incidentally, the possible reason why the band of DM-template in lane 4 is slightly thinner than those in other lanes (lanes 1-3) is the weakening Nb.BsmI falsely nicked DM-template similarly to normal restriction enzymes.
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