GWC Technologies

GWC
Technologies

505 S. Rosa Rd
Madison, WI 53719
USA

608.441.2726

GENOMICS
Genomics applications for GWC’s SPRimager®II system require no labelling of nucleic acids, and like other experiments, can involve molecules of any chemical composition. For example, you can monitor and quantify the binding of transcription factors to specific ss or dsDNA sequences on oligonucleotide arrays, enabling you to perform robust, direct comparisons of multiple DNA-protein interactions under identical conditions.

For detection of specific DNA targets at femotomolar concentration, we recommend the RNAseH system, which is unique in requiring neither amplification nor labelling of target DNA. Instead, DNA targets are detected on label-free RNA arrays, and signals are amplified via ribonuclease digestion. Based on the method of Goodrich et al, the method involves the following steps:

Mix target DNA sample with Ribonuclease H (RNase H) and buffer
Apply the mixture to an RNA oligonucleotide array prepared on SpotReady™ or SPRchip™ substrates
Monitor SPR signals for 15-120 minutes on the SPRimager®II at 30°: homologous RNA probes are degraded in proportion to the concentration of target DNA in the sample.

The mechanism of detection is loss of SPR signal due to degradation of RNA oligos by the RNase H, which degrades RNA only in the RNA-DNA duplex formed when complementary target DNA anneals.
The mechanism of amplification is recycling of the DNA targets and RNase H—once the RNA probe is degraded, the DNA target is released and can hybridize to another complementary RNA probe molecule; this hybrid duplex is again degraded by the RNase H, etc.

To learn more about using AmpliFast™, including how to apply the method to gene expression analysis, please contact .

Peer-reviewed experiments on genomics and development of the RNAseH method are cited on the publications section. Validated genomics applications for the SPRimager®II platform include:

*Patents pending