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PEMC-based method of measuring DNA hybridization at femtomolar concentration directly in human serum and in the presence of copious noncomplementary strands
Journal article   Peer reviewed

PEMC-based method of measuring DNA hybridization at femtomolar concentration directly in human serum and in the presence of copious noncomplementary strands

Kishan Rijal and Raj Mutharasan
Analytical chemistry (Washington), v 79(19), pp 7392-7400
01 Oct 2007
PMID: 17764156

Abstract

Base Sequence Biosensing Techniques DNA - blood DNA - chemistry Humans Kinetics Nucleic Acid Hybridization Oligonucleotide Probes Sensitivity and Specificity
Piezoelectric-excited, millimeter-sized cantilever (PEMC) sensors having high-mode resonance near 1 MHz are shown to exhibit mass change sensitivity of 1-300 ag/Hz. Gold-coated PEMC sensors immobilized with 15-mer single-stranded DNA (ssDNA) were exposed to 10-mer complementary strands at concentrations of 1 fM, 1 pM, and 1 microM, both separately and sequentially at 0.6 mL/min in a sample flow cell housing the sensor. Decrease in resonance frequency occurred as complementary strands hybridized to the immobilized probe DNA on the sensor surface. Hybridization in three background matrixes--buffer, buffer containing 10,000 times higher noncomplementary strands, and 50% human plasma--were successfully tested. Sensor hybridization responses to 1 fM, 1 pM, and 1 microM complementary strand were nearly the same in magnitude in all three matrixes, but the hybridization rates were different. In each case, the sensor detected the presence of 2 amol of complementary 10-mer strand. The extent of hybridization calculated from resonance frequency change did not decrease in serum. The findings suggest ssDNA can be detected at 2 amol without a sample preparation step and without the use of labeled reagents.

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Chemistry, Analytical
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