Microspotting streptavidin and double-stranded DNA arrays on gold for high-throughput studies of protein-DNA interactions by surface plasmon resonance microscopy

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dc.contributor.author Zareie, Hadi en_US
dc.contributor.author Shumaker-Parry, Jennifer en_US
dc.contributor.author Aebersold, Ruedi en_US
dc.contributor.author Campbell, Charles en_US
dc.date.accessioned 2009-06-26T04:10:47Z
dc.date.available 2009-06-26T04:10:47Z
dc.date.issued 2004 en_US
dc.identifier 2006006462 en_US
dc.identifier.citation Zareie Hadi et al. 2004, 'Microspotting streptavidin and double-stranded DNA arrays on gold for high-throughput studies of protein-DNA interactions by surface plasmon resonance microscopy', ACS Publications, vol. 76, no. 4, pp. 918-929. en_US
dc.identifier.issn 0003-2700 en_US
dc.identifier.other C1UNSUBMIT en_US
dc.identifier.uri http://hdl.handle.net/10453/515
dc.description.abstract We present two strategies for microspotting 10 ? 12 arrays of double-stranded DNAs (dsDNAs) onto a gold-coated glass slide for high-throughput studies of protein-DNA interactions by surface plasmon resonance (SPR) microscopy. Both methods use streptavidin (SA) as a linker layer between a biotin-containing mixed self-assembled monolayer (SAM) and biotinylated dsDNAs to produce arrays with high packing density. The primary mixed SAM is produced from biotin- and oligo(ethylene glycol)-terminated thiols bonded as thiolates onto the gold surface. In the first method, a robotic microspotter is used to deliver nanoliter droplets of dsDNA solution onto a uniform layer of this SA (~2 ? 1012 SA/cm2). SPR microscopy shows a density of (5-6) ? 1011 dsDNA/cm2 (0.2-0.3 dsDNA/SA) in the array elements. The second method uses instead a microspotted array of this SA linker layer, onto which the microspots of dsDNA are added with spatial registry. SPR microscopy before addition of the dsDNA shows a SA coverage of 2 ? 1012 SA/cm2 within the spots and a dsDNA density of 8.5 ? 3.5 ? 1011 dsDNA/cm2 (0.3-0.7 dsDNA/SA, depending on the length of dsDNA) after dsDNA spotting. We demonstrate the ability to simultaneously monitor protein binding with the SPR microscope in many 200-m spots with 1-s time resolution and sensitivity to <1 pg of protein. en_US
dc.publisher ACS Publications en_US
dc.relation.isbasedon http://dx.doi.org/10.1021/ac034964v en_US
dc.title Microspotting streptavidin and double-stranded DNA arrays on gold for high-throughput studies of protein-DNA interactions by surface plasmon resonance microscopy en_US
dc.parent Analytical Chemistry en_US
dc.journal.volume 76 en_US
dc.journal.number 4 en_US
dc.publocation Ohio, USA en_US
dc.identifier.startpage 918 en_US
dc.identifier.endpage 929 en_US
dc.cauo.name SCI.Physics and Advanced Materials en_US
dc.conference Verified OK en_US
dc.for 030100 en_US
dc.personcode 030414 en_US
dc.personcode 0000030208 en_US
dc.personcode 0000030209 en_US
dc.personcode 0000030210 en_US
dc.percentage 100 en_US
dc.classification.name Analytical Chemistry en_US
dc.classification.type FOR-08 en_US


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