New More Efficient Technique for the Recovery of DNA

Antonio Garcia (Inventor)

Research output: Patent

Abstract

The analysis of biological materials such as DNA typically involves the isolation and recovery of extremely small amounts of material. Quite often, the amount of material is so small that laboratory procedures such as polymerase chain reaction (PCR) must be employed so as to provide sufficient material for accurate analysis. Critical to the successful application of PCR is the efficient concentration and isolation of DNA. Accepted methods of concentrating DNA typically employ reagent systems using organic solvents for precipitation. Although the technique allows for the quantitative recovery of nanogram-levels of material, it does however, cause concerns related to analyst health and safety because potentially harmful solvents are used. Consequently, alternatives not relying upon organic solvents have been developed. These include UV radiation and ion exchange chromatography. Each, however, have functional limitations. Chromatographic methods tend to be time-consuming, equipment intensive, and are generally better suited for microgram quantities of DNA, while, UV methods tend to be prone to contamination since dried DNA and short-strand contaminants (e.g. 90% using aqueous elution buffers. This promising method is currently being tested in a variety of applications ranging from plasmid recovery to forensic recovery of mtDNA. The method is believed to uniquely benefit applications requiring the recovery of trace amounts of DNA of short sequence length due to the reagent's reversible affinity for the certain bases using mild conditions. Similarly, the method can improve PCR primer synthesis since unincorporated mononucleotides can be easily separated from synthesized oligonucleotides.
Original languageEnglish (US)
StatePublished - Jan 1 1900

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Recovery
Polymerase chain reaction
DNA
Organic solvents
Chromatography
Mitochondrial DNA
Oligonucleotides
Ultraviolet radiation
Biological materials
Ion exchange
Buffers
Plasmids
Contamination
Health
Impurities

Cite this

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title = "New More Efficient Technique for the Recovery of DNA",
abstract = "The analysis of biological materials such as DNA typically involves the isolation and recovery of extremely small amounts of material. Quite often, the amount of material is so small that laboratory procedures such as polymerase chain reaction (PCR) must be employed so as to provide sufficient material for accurate analysis. Critical to the successful application of PCR is the efficient concentration and isolation of DNA. Accepted methods of concentrating DNA typically employ reagent systems using organic solvents for precipitation. Although the technique allows for the quantitative recovery of nanogram-levels of material, it does however, cause concerns related to analyst health and safety because potentially harmful solvents are used. Consequently, alternatives not relying upon organic solvents have been developed. These include UV radiation and ion exchange chromatography. Each, however, have functional limitations. Chromatographic methods tend to be time-consuming, equipment intensive, and are generally better suited for microgram quantities of DNA, while, UV methods tend to be prone to contamination since dried DNA and short-strand contaminants (e.g. 90{\%} using aqueous elution buffers. This promising method is currently being tested in a variety of applications ranging from plasmid recovery to forensic recovery of mtDNA. The method is believed to uniquely benefit applications requiring the recovery of trace amounts of DNA of short sequence length due to the reagent's reversible affinity for the certain bases using mild conditions. Similarly, the method can improve PCR primer synthesis since unincorporated mononucleotides can be easily separated from synthesized oligonucleotides.",
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N2 - The analysis of biological materials such as DNA typically involves the isolation and recovery of extremely small amounts of material. Quite often, the amount of material is so small that laboratory procedures such as polymerase chain reaction (PCR) must be employed so as to provide sufficient material for accurate analysis. Critical to the successful application of PCR is the efficient concentration and isolation of DNA. Accepted methods of concentrating DNA typically employ reagent systems using organic solvents for precipitation. Although the technique allows for the quantitative recovery of nanogram-levels of material, it does however, cause concerns related to analyst health and safety because potentially harmful solvents are used. Consequently, alternatives not relying upon organic solvents have been developed. These include UV radiation and ion exchange chromatography. Each, however, have functional limitations. Chromatographic methods tend to be time-consuming, equipment intensive, and are generally better suited for microgram quantities of DNA, while, UV methods tend to be prone to contamination since dried DNA and short-strand contaminants (e.g. 90% using aqueous elution buffers. This promising method is currently being tested in a variety of applications ranging from plasmid recovery to forensic recovery of mtDNA. The method is believed to uniquely benefit applications requiring the recovery of trace amounts of DNA of short sequence length due to the reagent's reversible affinity for the certain bases using mild conditions. Similarly, the method can improve PCR primer synthesis since unincorporated mononucleotides can be easily separated from synthesized oligonucleotides.

AB - The analysis of biological materials such as DNA typically involves the isolation and recovery of extremely small amounts of material. Quite often, the amount of material is so small that laboratory procedures such as polymerase chain reaction (PCR) must be employed so as to provide sufficient material for accurate analysis. Critical to the successful application of PCR is the efficient concentration and isolation of DNA. Accepted methods of concentrating DNA typically employ reagent systems using organic solvents for precipitation. Although the technique allows for the quantitative recovery of nanogram-levels of material, it does however, cause concerns related to analyst health and safety because potentially harmful solvents are used. Consequently, alternatives not relying upon organic solvents have been developed. These include UV radiation and ion exchange chromatography. Each, however, have functional limitations. Chromatographic methods tend to be time-consuming, equipment intensive, and are generally better suited for microgram quantities of DNA, while, UV methods tend to be prone to contamination since dried DNA and short-strand contaminants (e.g. 90% using aqueous elution buffers. This promising method is currently being tested in a variety of applications ranging from plasmid recovery to forensic recovery of mtDNA. The method is believed to uniquely benefit applications requiring the recovery of trace amounts of DNA of short sequence length due to the reagent's reversible affinity for the certain bases using mild conditions. Similarly, the method can improve PCR primer synthesis since unincorporated mononucleotides can be easily separated from synthesized oligonucleotides.

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