Linked Life Data

19093760

Source:http://linkedlifedata.com/resource/pubmed/id/19093760

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2009-1-14
pubmed:abstractText
We describe in detail a new electrochemical DNA (E-DNA) sensing platform based on target-induced conformation changes in an electrode-bound DNA pseudoknot. The pseudoknot, a DNA structure containing two stem-loops in which the first stem's loop forms part of the second stem, is modified with a methylene blue redox tag at its 3' terminus and covalently attached to a gold electrode via the 5' terminus. In the absence of a target, the structure of the pseudoknot probe minimizes collisions between the redox tag and the electrode, thus reducing faradaic current. Target binding disrupts the pseudoknot structure, liberating a flexible, single-stranded element that can strike the electrode and efficiently transfer electrons. In this article we report further characterization and optimization of this new E-DNA architecture. We find that optimal signaling is obtained at an intermediate probe density ( approximately 1.8 x 10(13) molecules/cm(2) apparent density), which presumably represents a balance between steric and electrostatic blocking at high probe densities and increased background currents arising from transfer from the pseudoknot probe at lower densities. We also find that optimal 3' stem length, which appears to be 7 base pairs, represents a balance between pseudoknot structural stability and target affinity. Finally, a 3' loop comprised of poly(A) exhibits better mismatch discrimination than the equivalent poly(T) loop, but at the cost of decreased gain. Optimization over this parameter space significantly improves the signaling of the pseudoknot-based E-DNA architecture, leading to the ability to sensitively and specifically detect DNA targets even when challenged in complex, multicomponent samples such as blood serum.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-11457381, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-11859188, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-12553781, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-12824337, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-14588003, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-14616004, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-14640732, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-15339145, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-15649056, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-15749017, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-16906710, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-17129062, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-17488132, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-17850085, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-18007622, http://linkedlifedata.com/resource/pubmed/commentcorrection/19093760-2720781
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
1520-6882
pubmed:author
pubmed:issnType
Electronic
pubmed:day
15
pubmed:volume
81
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
656-61
pubmed:dateRevised
2010-12-17
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Optimization of a reusable, DNA pseudoknot-based electrochemical sensor for sequence-specific DNA detection in blood serum.
pubmed:affiliation
Department of Chemical Engineering, Institute for Polymers and Organic Solids, University of California, Santa Barbara, California 93106, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, N.I.H., Extramural