DNA in the presence of gellan
First Claim
1. A method for enzymatically amplifying a target nucleic acid or a fragment thereof, comprising (a) providing a nucleic acid amplification reaction mixture having a water-based liquid phase and a gellan gel matrix phase, wherein the liquid phase comprises a target nucleic acid and is entrapped in the gel matrix phase;
- and (b) subjecting the reaction mixture to conditions suitable for amplifying the target nucleic acid or a fragment thereof, whereby the target nucleic acid or the fragment thereof is amplified.
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Abstract
A method is provided for nucleic acid amplification with enhanced sensitivity. The method for enhanced sensitivity involves carrying out the amplification reaction in the presence of gellan. For instance, the method allows for the production of detectible amounts of PCR amplified DNA from at least 10 fold fewer target molecules than a comparable PCR reaction in absence of gellan.
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Citations
16 Claims
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1. A method for enzymatically amplifying a target nucleic acid or a fragment thereof, comprising
(a) providing a nucleic acid amplification reaction mixture having a water-based liquid phase and a gellan gel matrix phase, wherein the liquid phase comprises a target nucleic acid and is entrapped in the gel matrix phase; - and
(b) subjecting the reaction mixture to conditions suitable for amplifying the target nucleic acid or a fragment thereof, whereby the target nucleic acid or the fragment thereof is amplified. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method for enzymatically amplifying a target nucleic acid or a fragment thereof, comprising
(a) providing a nucleic acid amplification reaction mixture that comprises a target nucleic acid, and gellan at a concentration above 0.005 wt %; - and
(b) subjecting the reaction mixture to conditions suitable for amplifying the target nucleic acid or a fragment thereof, whereby the target nucleic acid or the fragment thereof is amplified. - View Dependent Claims (10, 11, 12, 13, 14)
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15. In a method for enzymatically amplifying nucleic acid, the improvement comprising performing the enzymatic amplification in the presence of at least 0.005 wt % gellan or gellan fragments.
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16. A composition suitable for use in nucleic acid amplification comprising water, gellan at a concentration above 0.005 wt % based on the weight of water, a DNA polymerase, dNTPs, and a target nucleic acid.
Specification