Journal
MICROCHIMICA ACTA
Volume 177, Issue 3-4, Pages 435-441Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-012-0799-0
Keywords
Surface plasmon resonance (SPR); Taq polymerase; Mismatch; Caspase-3
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Funding
- National Natural Science Foundation of China [31070772]
- Doctoral Program of Higher Education [200901011110136]
- Science and Technology Programs of Zhejiang Province [2011 C37029]
- Science and Technology Programs of Suzhou [ZXG0920]
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We demonstrate that base mismatches of caspase-3 DNA sequences can be detected by surface plasmon resonance (SPR) following signal amplification by polymerase from (). The concentration of magnesium ions and the respective dNTPs for polymerase binding to the oligonucleotides on the sensing surface were optimized. polymerase binds to double-stranded DNA that is self-assembled on the gold surface of the biosensor to induce an SPR signal. Experiments are presented on the effect of Mg(II) and dNTP concentrations on the activity of the polymerase on the sensing surface. The detection limits are 50 pM, 0.1 nM, 0.7 nM, 7 nM, and 20 nM for correctly matched, single-base mismatched, two-base mismatched, three-base mismatched and four-base mismatched DNA of caspase-3, respectively. This is attributed to the optimized experimental conditions, with samples containing 2 mu M of Mg(II) and 0.3 mM of dNTP.
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