A method to create DNA copies of specific genetic material using primers and enzymes, enabling easier detection, amplification, cloning, and sequencing of DNA and RNA.
Background
Advancements in molecular biology have underscored the critical need for efficient and accurate methods to prepare DNA copies of target polynucleotides. Techniques such as PCR amplification, cloning, and sequence determination are fundamental for applications ranging from medical diagnostics to genetic research.
As the demand for high-throughput and precise genetic analysis grows, the ability to generate reliable DNA copies becomes increasingly essential. Enhanced methodologies in DNA preparation facilitate better detection, enable more robust amplification processes, and improve the overall accuracy of genetic sequencing, thereby driving forward innovations in biotechnology and personalized medicine. .
However, current approaches to DNA copy preparation often encounter significant challenges that hinder their effectiveness. Traditional methods may suffer from low efficiency in primer extension, leading to incomplete or inaccurate DNA synthesis. Additionally, existing template switching techniques can be limited by the fidelity of reverse transcriptases, resulting in errors during the copying process. These limitations can impede the reliability of downstream applications such as PCR amplification and cloning, reducing the overall quality and reproducibility of genetic analyses. Furthermore, the complexity and time-consuming nature of existing protocols can restrict their scalability and accessibility, posing obstacles to widespread adoption in both research and clinical settings.
Technology description
The technology involves a method for creating a DNA copy of a target polynucleotide through template switching. It begins by combining a double-stranded template/primer substrate, which consists of a DNA primer oligonucleotide paired with a complementary template strand, with the target polynucleotide in a reaction medium. A non-retroviral reverse transcriptase is then added to extend the DNA primer from its 3′ end, resulting in the synthesis of a DNA copy that includes a complementary strand to the target polynucleotide. Additionally, the method outlines techniques for incorporating nucleotides into the double-stranded template/primer substrate. This process is versatile, enabling applications such as detection, PCR amplification, cloning, and the sequencing of RNA and DNA.
What sets this technology apart is its use of non-retroviral reverse transcriptase for template switching, enhancing the precision and efficiency of DNA synthesis from target polynucleotides. Unlike traditional methods, this approach allows for more accurate and reliable amplification and cloning, reducing errors during the copying process.
The ability to effectively synthesize complementary DNA strands facilitates a wide range of molecular biology techniques, making it highly adaptable for various applications in genetic research and diagnostics. Its streamlined process not only improves the quality of DNA copies but also expands the potential for more sophisticated genetic analyses and manipulations.
Benefits
- Enables efficient preparation of DNA copies from target polynucleotides
- Facilitates detection, PCR amplification, cloning, and sequencing of nucleic acids
- Utilizes non-retroviral reverse transcriptase for reliable DNA synthesis
Commercial applications
- Genomic sequencing services
- PCR amplification kits
- DNA cloning reagents
- RNA sequencing platforms
- Diagnostic assay development
Opportunity
The University of Texas at Austin is seeking a commercial partner to license and/or manufacture this technology.
Intellectual property
Issued patents: US 9,012,183; CA 2,827,948; JP 6068365; CN ZL2012800095894.1; EP 2678430; IL 228042; KR 10-1911969; FR 2678430; GB 2678430; DE 602012045063; HK HK1203218A1
