Method for the Selective Enrichment of Double-Stranded Dna from Nucleic Acid Mixtures

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This patent discloses a method for selectively enriching double-stranded DNA, especially supercoiled plasmid DNA, from mixtures containing both single-stranded and double-stranded nucleic acids. The method involves reversible denaturation using alkaline or heat treatment, followed by extraction in an aqueous two-phase system (containing specific buffers and polyethylene glycol), which separates the single-stranded nucleic acids from the desired double-stranded DNA. The process is suitable for both small- and large-scale purification needs.

Use CasesContent extracted from patent full text and abstract with AI.

• Preparation of high-purity supercoiled plasmid DNA for gene therapy and DNA vaccination
• Production of DNA for genetic research and molecular biology experiments
• Manufacture of clinical-grade plasmid DNA for pharmaceutical and biotechnology companies
• DNA sample preparation for high-throughput or diagnostic applications
• Automated large-scale plasmid DNA purification in industrial settings
• Preparation of clean DNA samples in academic or commercial research laboratories

BenefitsContent extracted from patent full text and abstract with AI.

• Highly efficient and selective removal of single-stranded DNA, RNA, and open-circle plasmid DNA from supercoiled plasmid DNA
• Lower cost and faster processing compared to conventional chromatographic methods
• Scalable from manual kit-level to industrial production scale
• Amenable to automation for high-throughput or routine manufacturing
• Uses non-toxic, easily removable reagents, making it environmentally friendly
• Significantly improves purity and yield of target DNA, meeting stringent clinical and regulatory requirements
• Reduces the number of purification steps needed, lowering risks and overall costs

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Patent Abstract

Method for removing single-stranded nucleic acid (sNA) from double-stranded nucleic acid (dNA) by selective, reversible denaturation. Method for removing single-stranded nucleic acid (sNA) from double-stranded nucleic acid (dNA) comprises: (A) providing a mixture containing completely and/or partially double-stranded nucleic acid, optionally also sNA; (B) resuspension in dilute, aqueous buffer of low ionic strength and low buffering capacity; (C) adjusting concentration of the mixture so as to cause reversible denaturation of one or more particular dNA, while other nucleic acids is/are denatured irreversibly; (D) adding more buffer and a polymer; (E) incubating for long enough to form an aqueous 2-phase system with upper and lower phases; and (F) removing the sNA-containing upper (or intermediate) phase and collecting dNA from the lower phase.