Single-Step Method to Fragment and Tag Both Ends of DNA Molecules with Arbitrary Nucleotide Sequences

Web Published:
6/9/2017
Description:

One-step method to fragment and tag both ends of DNA for Next Generation Sequencing (NGS)

Princeton Docket # 17-3313

 

 

Researchers in the Lewis-Sigler Institute for Integrative Genomics at Princeton University have developed a new simplified method to fragment and tag both ends of DNA molecules with arbitrary nucleotide sequences for Next Generation Sequencing as well as other high-throughput DNA processing.

 

This invention combines all reagents in a single step to fragment and tag both ends of DNA molecules with desired nucleotide sequences with optional PCR amplification, while all other existing NGS library preparation methods involve multiple steps which use different reagents. 

 

This invention not only reduces the hands-on sample processing time, but also greatly facilitates the high-throughput compartmentalized DNA processing, such as water-in-oil emulsion droplets, which are not amenable to using different reagents in multiple steps.  This invention can potentially be used in other types of compartmentalized micro-reactors and nano-biotechnologies.

 

Applications        

•       DNA library preparation for Next-Generation Sequencing

•       Emulsion droplet based DNA processing

•       Nano-biotechnologies and other micro-reactors on DNA

 

Advantages        

•       Simplest 1-step method

•       Single reaction mixture including all reagents and DNA sample

•       Reaction controlled by a series of incubation temperatures

•       Reduced sample processing time

•       Optional final PCR amplification

 

Stage of development

 

The method has been validated on genomic DNA samples to produce sequencing libraries compatible with the Illumina sequencers. The produced libraries have been sequenced on the Illumina HiSeq 2500 sequencer in our Genomics Core Facility, and preliminary bioinformatics analysis indicated these data successfully covered the target genome. 

 

Inventor

Wei Wang, Ph.D. is the Director of the Genomics Core Facility at Princeton University. In addition to supervising the core laboratory to provide genomics services using publicly available protocols and reagents, he focuses on developing novel genomics methods, especially in Next-Generation Sequencing (NGS), to address the unmet needs in life science research.  Currently he is working on single-cell RNA and DNA amplification and library preparation, simple and efficient sample processing methods for microfluidics and/or emulsion droplet-based genomics assays.

Intellectual Property & Development status

Patent protection is pending.

Princeton is currently seeking commercial partners for the further development and commercialization of this opportunity.

Contact

 

Laurie Tzodikov

Princeton University Office of Technology Licensing • (609) 258-7256• tzodikov@princeton.edu

 

 

 

 

 

Patent Information:
For Information, Contact:
Chris Wright
Licensing Associate
Princeton University
cw20@princeton.edu
Inventors:
Wei Wang
Keywords: