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Gothami Padmabandu Illumina Inc, United States of America |
Abstract
Rreal-time quantitative PCR (RT-qPCR) has been widely used in the analysis of gene expression for decades. Currently many probe based chemistries have been utilized for RT-qPCR analysis. NūPCR reagents constitute a novel probe-based technology that utilizes a unique NūZyme chemistry. NūZymes are multipart catalytic enzymes that recognize and assemble on target nucleic acid sequences forming a catalytic complex. This three part probe consists of two PartZyme oligos and a universal substrate oligo containing a 5’ flourophore and a 3’ quencher that binds to the partzyme oligo. Once the target is amplified via PCR, the two partzymes bind to the specific target sequence forming the NūZyme complex. The enzymatic activity of the NūZyme cleaves the fluorescently labeled universal substrate producing a signal that can be detected by a real-time PCR instrument.
The high sensitivity, specificity and ease of designing duplexed assays will be presented. Different applications of NūPCR such as a highly multiplexed DNA quantitation assay for forensic samples will be discussed.
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