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Probing concentrati...
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Frykholm, Karolin,1977Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU),University of Gothenburg
(författare)
Probing concentration-dependent behavior of DNA-binding proteins on a single-molecule level illustrated by Rad51
- Artikel/kapitelEngelska2013
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LIBRIS-ID:oai:gup.ub.gu.se/189119
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https://gup.ub.gu.se/publication/189119URI
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https://doi.org/10.1016/j.ab.2013.08.023DOI
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https://lup.lub.lu.se/record/4196484URI
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https://research.chalmers.se/publication/189119URI
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-212824URI
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Low throughput is an inherent problem associated with most single-molecule biophysical techniques. We have developed a versatile tool for high-throughput analysis of DNA and DNA-binding molecules by combining microfluidic and dense DNA arrays. We use an easy-to-process microfluidic flow channel system in which dense DNA arrays are prepared for simultaneous imaging of large amounts of DNA molecules with single-molecule resolution. The Y-shaped microfluidic design, where the two inlet channels can be controlled separately and precisely, enables the creation of a concentration gradient across the microfluidic channel as well as rapid and repeated addition and removal of substances from the measurement region. A DNA array stained with the fluorescent DNA-binding dye YOYO-1 in a gradient manner illustrates the method and serves as a proof of concept. We have applied the method to studies of the repair protein Rad51 and could directly probe the concentration-dependent DNA-binding behavior of human Rad51 (HsRad51). In the low-concentration regime used (100 nM HsRad51 and below), we detected binding to double-stranded DNA (dsDNA) without positive cooperativity. (C) 2013 Elsevier Inc. All rights reserved.
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Biuppslag (personer, institutioner, konferenser, titlar ...)
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Freitag, CamillaGothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU),University of Gothenburg
(författare)
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Persson, Fredrik,1979Uppsala universitet,Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU),University of Gothenburg,Beräknings- och systembiologi(Swepub:uu)frepe558
(författare)
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Tegenfeldt, Jonas O.Lund University,Lunds universitet,Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU),Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH,University of Gothenburg(Swepub:lu)ftf-jte
(författare)
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Graneli, Annette,1973Gothenburg University,Göteborgs universitet,Institutionen för fysik (GU),Department of Physics (GU),University of Gothenburg(Swepub:gu)xgrane
(författare)
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Göteborgs universitetInstitutionen för fysik (GU)
(creator_code:org_t)
Sammanhörande titlar
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Ingår i:Analytical Biochemistry: Elsevier BV443:2, s. 261-2680003-26971096-0309
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