Enzymatically amplified surface plasmon resonance imaging method using RNase H and RNA microarrays for the ultrasensitive detection of nucleic acids.
A novel surface enzymatic amplification method that utilizes RNA microarrays in conjunction with the enzyme RNase H is developed for the ultrasensitve detection and analysis of target DNA molecules. The enzyme RNase H is shown to selectively and repeatedly destroy RNA from RNA-DNA heteroduplexes on gold surfaces; when used in conjunction with the label-free technique of surface plasmon resonance imaging, multiple DNA targets can be detected at a concentration of 10 fM on a single chip. In addition, this method is utilized for the sequence-specific detection of the TSPY gene in both purified and unpurified PCR products. Finally, in a series of kinetics measurements, the initial rate of hydrolysis is shown to depend directly on the surface concentration of DNA-RNA heteroduplexes.
Goodrich
Terry T
TT
Department of Chemistry, University of California-Irvine, Irvine, California 92697, USA.
Lee
Hye Jin
HJ
Corn
Robert M
RM
D001483:Base Sequence; D009692:Nucleic Acid Heteroduplexes; D009696:Nucleic Acids / Q000032:analysis*; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000737:chemistry*; D016914:Ribonuclease H / Q000737:chemistry*; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D009692:Nucleic Acid Heteroduplexes; D009696:Nucleic Acids; D012313:RNA; D016914:Ribonuclease H