Photoinduced Electron Transfer Fluorescent Primer for Nucleic Acid Amplification


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.

Potential Commercial Applications: Competitive Advantages:
  • Efficient fluorescence-labeling of oligonucleotides
  • Quantitative methods
  • Pyro-sequencing
  • Basic laboratory research
 
  • 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


Development Stage:
In vitro data available

Inventors:

Jothikumar Narayanan (CDC)  ➽ more inventions...

Vincent Hill (CDC)  ➽ more inventions...

Brian Holloway (CDC)  ➽ more inventions...


Intellectual Property:
U.S. Pat: 9,260,746 issued 2016-02-16
US Application No. 60/989,768
PCT Application No. PCT/US2008/084347
US Application No. 12/743,607
Various international filings pending

Publications:
Jothikumar N, Hill VR. PMID 23727382

Licensing Contact:
Tara Kirby, Ph.D.
Email: tara.kirby@nih.gov
Phone: 240-669-5128

OTT Reference No: E-292-2013/0
Updated: Feb 17, 2016