Quadruplex-Based Technology for Mono-Adapter DNA Library Preparation and Isothermal Clone Generation for Next Generation Sequencing (NGS)

A technique for the isothermal amplification of DNA libraries for a more efficient and robust sequencing method.

The Need

Identification of a particular nucleic acid is desirable in the use of diagnostic and forensic applications. However, often the target nucleic acid sequence may only be a small portion of the DNA or RNA in question so that amplification is needed before sufficient quantities are available for identification. Polymerase Chain Reaction (PCR) amplification, the current gold standard, is used to amplify the target sequence. Drawbacks of PCR processes and probe technologies include competition between primer binding and self-annealing, high frequency temperature cycling, and cost. An alternative technology that can reduce complications and expenses and shorten workflow would improve identification of nucleic acids for a variety of commercial applications.

The Technology

The Ohio State University researchers led by Dr. Besik Kankia developed a quadruplex priming amplification (QPA) technology to improve amplification and identification of nucleic acids. They refined the technique to allow for isothermal amplification of a DNA library. To reduce waste from dissimilar ends (A-B) and shorten sequencing workflow resulting from one direction sequencing, they have improved the method to make pair-end sequencing possible by having the same adapter at both ends. This novel technology will greatly improve nucleic acid amplification by skipping the enrichment step, using very little genomic material, and using a mono-adapter approach to shorten sequencing times.

Commercial Applications

  • Pharmaceutical development
  • Forensics
  • Genotyping

Benefits/Advantages

  • Reduces waste of samples by using mono-adapters
  • Shortens sequencing workflow times
  • Decreases cost
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