Eric Tucker - Chemical amplification for in-gel DNA detection

Document created by Eric Tucker on Aug 22, 2014
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  Publication Details (including relevant citation   information): He, Peng, Tucker, Eric Z., Gorman,   Christopher B., He, Lin, Anal. Methods,   2011, 3 (11), pp 2463-2468

  Abstract: We report the use of reversible   addition-fragmentation chain transfer (RAFT) polymerization as a   highly efficient chemical amplification means to direct   visualization of DNA in porous polyacrylamide gel. It is the   first time that a dynamic polymer growth on the surface of soft   medium is used in signal amplification for DNA detection. In the   proof-of-concept experiment, a thin acrylamide gel on a glass   microscope slide formed a thin layer of uniformly crosslinked   network with porous structures. Oligonucleotides of different   sequences were entrapped within the gel at separate spots.   Hybridization of complementary DNA detection probes introduced   chain transfer agents (CTAs) into the gel via preconjugation to   the probes. Surface-initiated polymer growth was prompted on the   gel surface and the growth of polymer brushes at the spot where   DNA hybridization occurred was monitored using infrared   spectroscopy and atomic force microscopy. Visible change in the   texture of the porous gel occurred after polymer growth, which   offered an attractive detection alternative for in-gel DNA   analysis. Compared to the results from traditional ethidium   bromide staining, better detection sensitivity and specificity   were achieved.

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