APPLICATIONS OF TECHNOLOGY:
- Forensic investigation
- Clinical research
- Medical diagnostics
- National security
- Biosecurity protection
- High volume, low biomass samples
- Greater efficiency of extracting trace DNA from field samples
- Higher sensitivity for trace amounts of DNA
- High selectivity for specific DNA of interest in a complex sample
- Improved purity of DNA extracts
- Increased compatibility with a broad range of downstream analyses
- Versatility for extraction of minute amounts of DNA in a large sample
Youn-Hi Woo of Berkeley Lab’s Physical Biosciences Division has invented a technology to extract and isolate DNA from complex field samples that improves the efficiency of the process and the purity of the sample. This technology widens the possibilities for downstream analysis techniques due to the resulting higher degree of purity of the DNA sample and the high sensitivity to minute amounts of DNA in the sample.
Using the DNA-specific enzyme methyltransferase (Mtase) and a “click” chemistry process, Woo’s technique is a new platform that addresses these challenges. While current methods detect DNA to the nanogram range, the Mtase method has shown high sensitivity and selectivity as low as the sub-femtogram range of DNA. Preliminary results in the lab show that the technology can detect as little as one molecule of DNA in a sample. The extracted DNA is subsequently captured through covalent bond to a functionalized surface using click chemistry. Various surface matrices can be used: glass, plastic tubes, plates, and magnetic or polymer beads. This allows a far more vigorous scrubbing process to remove impurities and carbohydrates, without compromising or losing the DNA.
This process guarantees that DNA will be the only final product rather than DNA in a mixture of other substances and impurities. Woo’s technology can be applied to any volume, down to trace amounts, making it an alternative for larger volume samples. Because of the purity of the sample obtained, the extract can be combined with a broader range of downstream analyses than a sample obtained using current methods.
Woo’s method holds great promise for applications where extraction of minute amounts of DNA plays a critical role, such as in basic and applied molecular biology research, bioforensics, biosecurity sciences, and state-of-the-art detection methods. Where the sample size is very large, Woo’s technique will enable the detection of minute amounts of DNA.
DNA has become an important player in providing clues for research, forensic investigations, and diagnostics, but common methods for extracting it have created problems for “downstream” analyses. Samples can contain impurities that hamper subsequent usefulness. There is an urgent need in research for reliable new methods to extract DNA in various conditions and at higher sensitivity. The current limit on sample sizes is an additional inhibiting factor in many investigations.
STATUS: Patent pending. Available for licensing or collaborative research.
FOR MORE INFORMATION:
Artyukhin, A.B., Woo, Y.-H. “DNA extraction method with improved efficiency and specificity using DNA methyltransferase and ‘click’ chemistry,” Analytical Biochemistry, 425, June 15, 2012, 2, 169-174.
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REFERENCE NUMBER: IB-2780