Abstract
ABSTRACT DNA/mRNA analysis and genetic mutation detection are important in biomedical sciences, clinical diagnosis, and environmental monitoring. In this study, a femtoliter microwell array system has been developed for ultrasensitive DNA/mRNA detection using molecular beacon (MB) DNA probes. The microwell array system, with laser-induced fluorescence imaging, can be used to achieve a detection limit of 9 Rhodamin 6G molecules in 28 fL (28 × 10−15 L) wells in this array. Hybridization kinetics of the MBs and their complementary DNA targets has been monitored with a concentration detection limit of 3.0 nM. As few as 50 target DNA copies can be monitored in each well. The specificity of MB and the effect of DNA target concentration on the MB hybridization kinetics have been investigated in femtoliter wells. Analysis of specific rat γ-actin mRNA sequence amplified by polymerase chain reaction (PCR) and simultaneous measurement of multiple analytes have also been studied with this microwell array system. As the volume in each well is extremely small, this array system will be highly useful for single cell gene profiling and for multiple gene determination in disease diagnosis.
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