Combination of fluorescence color and melting temperature as a two-dimensional label for homogeneous multiplex PCR detection.

ABSTRACT: Multiplex analytical systems that allow detection of multiple nucleic acid targets in one assay can provide rapid characterization of a sample while still saving cost and resources. However, few systems have proven to offer a solution for mid-plex (e.g. 10- to 50-plex) analysis that is high throughput and cost effective. Here we describe the combined use of fluorescence color and melting temperature (Tm) as a virtual 2D label that enables homogenous detection of one order of magnitude more targets than current strategies on real-time polymerase chain reaction platform. The target was first hybridized with a pair of ligation oligonucleotides, one of which harbored an artificial sequence that had a unique Tm when hybridized with a reporter fluorogenic probe. The ligated products were then am

SUBMITTER: Liao Y 

PROVIDER: S-EPMC3627564 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature