Technology Bundle ID: TAB-2745

Photoinduced Electron Transfer Fluorescent Primer for Nucleic Acid Amplification

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Primary Inventors: 
Jothikumar Narayanan (CDC)
Co-Inventors: 
Brian Holloway (CDC), Vincent Hill (CDC)
Development Stage: 
Pre-Clinical (in vitro)
Development Status: 

In vitro data available

Institute or Center: 
CDC

CDC scientists have developed a rapid and cost-efficient method for generating fluorescently labeled primers for PCR and real-time PCR. At present, fluorescent primers are useful for detecting and identifying microbes and specific nucleic acid sequences, amplifying nucleic acids for pyro-sequencing, determining the levels of gene expression, and many other uses. However, problems exist with current techniques used to create fluorescent primers. For one, labeling is not one hundred percent efficient, leading to inaccurate results. Further, it is expensive and time consuming for researchers to make and label their own unique primers. This technology allows for the creation of custom primers in which fluorescent dye attaches to all oligomers.

This technology employs photoinduced electron transfer (PET) nucleic acid molecules that can be used detect and amplify target nucleic acid molecules. PET tags are attached to the 5'-end of a target-specific oligo for fluorescent labeling of the primer. PET tag activity can be quenched by at least two consecutive guanosines (G-G) within the tag sequence and activity is un-quenched when the PET tag hybridizes with its complementary nucleic acid molecule.

Applications:
  • Efficient fluorescence-labeling of oligonucleotides
  • Quantitative methods
  • Pyro-sequencing
  • Basic laboratory research
Advantages:
  • Avoids aberrant quantitative data generation resulting from inefficient fluorescent labeling reactions
  • Allows for multiplex reactions
  • Cost-efficient for time, sample preservation and cost of analysis
  • Method can readily be used as part of an oligo-labeling kit
  • No need for HPLC purification
  • Does not require a quencher dye

Patents

PCT Application PCT/US2008/084347
Filed on 2008-11-21
US Application 15/043,713
Filed on 2016-02-15
US Application 60/989,768
Filed on 2007-11-21
US Pat 9,260,746

Issued 2016-02-16

Various international filings pending

Publications

Jothikumar N, Hill VR.
PMID 23727382

Updated

Mar 9, 2020

Data Source: 
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