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Sökning: onr:"swepub:oai:lup.lub.lu.se:8687b89d-baef-47cb-ba48-84cd8f0131f6" > Single-molecule Det...

Single-molecule Detection and Mismatch Discrimination of Unlabeled DNA Targets

Gunnarsson, Anders (författare)
Lund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
Jönsson, Peter (författare)
Lund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
Marie, Rodolphe (författare)
Lund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
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Tegenfeldt, Jonas (författare)
Lund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
Höök, Fredrik (författare)
Lund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
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 (creator_code:org_t)
The American Chemical Society (ACS), 2008
2008
Engelska.
Ingår i: Nano Letters. - : The American Chemical Society (ACS). - 1530-6992. ; 8:1, s. 183-188
  • Tidskriftsartikel (refereegranskat)
Abstract Ämnesord
Stäng  
  • We report on a single-molecule readout scheme on total internal reflection fluorescence microscopy (TIRFM) demonstrating a detection limit in the low fM regime for short (30-mer) unlabeled DNA strands. Detection of unlabeled DNA targets is accomplished by letting them mediate the binding of suspended fluorescently labeled DNA-modified small unilamellar vesicles (Ø approximately 100 nm) to a DNA-modified substrate. On top of rapid and sensitive detection, the technique is also shown capable of extracting kinetics data from statistics of the residence time of the binding reaction in equilibrium, that is, without following neither the rate of binding upon injection nor release upon rinsing. The potential of this feature is demonstrated by discriminating a single mismatch from a fully complementary sequence. The success of the method is critically dependent on a surface modification that provides sufficiently low background. This was achieved through self-assembly of a biotinylated copolymer, Poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG) on a silicon dioxide surface, followed by subsequent addition of streptavidin and biotinylated DNA. The proposed detection scheme is particularly appealing due to the simplicity of the sensor, which relies on self-assembly principles and conventional TIRFM. Therefore, we foresee a great potential of the concept to serve as an important component in future multiplexed sensing schemes. This holds in particular true in cases when information about binding kinetics is valuable, such as in single nucleotide polymorphism diagnostics.

Ämnesord

TEKNIK OCH TEKNOLOGIER  -- Nanoteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Nano Technology (hsv//eng)
TEKNIK OCH TEKNOLOGIER  -- Nanoteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Nano-technology (hsv//eng)

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Av författaren/redakt...
Gunnarsson, Ande ...
Jönsson, Peter
Marie, Rodolphe
Tegenfeldt, Jona ...
Höök, Fredrik
Om ämnet
TEKNIK OCH TEKNOLOGIER
TEKNIK OCH TEKNO ...
och Nanoteknik
TEKNIK OCH TEKNOLOGIER
TEKNIK OCH TEKNO ...
och Nanoteknik
Artiklar i publikationen
Nano Letters
Av lärosätet
Lunds universitet

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