Direct detection of genomic DNA by enzymatically amplified SPR imaging measurements of RNA microarrays.
A novel surface enzymatic reaction scheme that amplifies the optical response of RNA microarrays to the binding of complementary DNA is developed for the direct detection and analysis of genomic DNA. The enzyme RNase H is shown to selectively and repeatedly destroy RNA from DNA-RNA heteroduplexes on gold surfaces; when used in conjunction with the label-free technique of surface plasmon resonance (SPR) imaging, DNA oligonucleotides can be detected at a concentration of 1 fM. This enzymatically amplified SPR imaging methodology is then utilized to detect and identify the presence of the TSPY gene in human genomic DNA without PCR amplification.
Goodrich
Terry T
TT
Department of Chemistry, University of Wisconsin, Madison, Wisconsin, USA.
Lee
Hye Jin
HJ
Corn
Robert M
RM
D004247:DNA / Q000032:analysis* / Q000737:chemistry; D020869:Gene Expression Profiling; D015894:Genome, Human; D015698:Genomic Library; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000032:analysis / Q000378:metabolism; D015347:RNA Probes / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000379:methods*
D015347:RNA Probes; D012313:RNA; D004247:DNA; D016914:Ribonuclease H