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Sensitive and inexp...
Sensitive and inexpensive digital DNA analysis by microfluidic enrichment of rolling circle amplified single-molecules
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- Kühnemund, Malte (författare)
- Uppsala universitet,Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Uppsala University, Sweden,Science for Life Laboratory, SciLifeLab,Institutionen för immunologi, genetik och patologi,Stockholm Univ, Dept Biophys & Biochem, Sci Life Lab, SE-17165 Solna, Sweden.
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- Hernández-Neuta, Iván, 1986- (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biophys & Biochem, Sci Life Lab, SE-17165 Solna, Sweden.
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- Sharif, Mohd Istiaq (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Stockholm Univ, Dept Biophys & Biochem, Sci Life Lab, SE-17165 Solna, Sweden.
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- Cornaglia, Matteo (författare)
- Ecole Polytech Fed Lausanne, Lab Microsyst, CH-1015 Lausanne, Switzerland.
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- Gijs, Martin A. M. (författare)
- Ecole Polytech Fed Lausanne, Lab Microsyst, CH-1015 Lausanne, Switzerland.
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- Nilsson, Mats (författare)
- Uppsala universitet,Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),Uppsala University, Sweden,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab,Stockholm Univ, Dept Biophys & Biochem, Sci Life Lab, SE-17165 Solna, Sweden.
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(creator_code:org_t)
- 2017-01-10
- 2017
- Engelska.
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Ingår i: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 45:8
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Single molecule quantification assays provide the ultimate sensitivity and precision for molecular analysis. However, most digital analysis techniques, i.e. droplet PCR, require sophisticated and expensive instrumentation for molecule compartmentalization, amplification and analysis. Rolling circle amplification (RCA) provides a simpler means for digital analysis. Nevertheless, the sensitivity of RCA assays has until now been limited by inefficient detection methods. We have developed a simple microfluidic strategy for enrichment of RCA products into a single field of view of a low magnification fluorescent sensor, enabling ultra-sensitive digital quantification of nucleic acids over a dynamic range from 1.2 aM to 190 fM. We prove the broad applicability of our analysis platform by demonstrating 5-plex detection of as little as ∼1 pg (∼300 genome copies) of pathogenic DNA with simultaneous antibiotic resistance marker detection, and the analysis of rare oncogene mutations. Our method is simpler, more cost-effective and faster than other digital analysis techniques and provides the means to implement digital analysis in any laboratory equipped with a standard fluorescent microscope.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Microbiology in the medical area (hsv//eng)
Nyckelord
- Biochemistry
- biokemi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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