Electrochemical denaturation of double-stranded nucleic acid
First Claim
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1. A process for denaturing double-stranded nucleic acid comprising the steps of:
- applying a voltage to a solution containing said nucleic acid with an electrode; and
converting at least a proportion of said nucleic acid to a wholly or partially single stranded form so as to produce single-stranded nucleic acid in said solution not bound to said electrode, which single-stranded nucleic acid is capable of renaturation to double-stranded form.
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Abstract
A process is described for denaturing native double-stranded nucleic acid material into its individual strands in an electrochemical cell. The process disclosed is an electrical treatment of the nucleic acid with a voltage applied to the nucleic acid material by an electrode. The process may also employ a promotor compound such as methyl viologen to speed denaturation. The process may be used in the detection of nucleic acid by hybridizing with a labelled probe or in the amplification of DNA by a polymerase chain reaction or ligase chain reaction.
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Citations
30 Claims
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1. A process for denaturing double-stranded nucleic acid comprising the steps of:
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applying a voltage to a solution containing said nucleic acid with an electrode; and converting at least a proportion of said nucleic acid to a wholly or partially single stranded form so as to produce single-stranded nucleic acid in said solution not bound to said electrode, which single-stranded nucleic acid is capable of renaturation to double-stranded form. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A process for amplifying a target sequence of nucleic acid comprising cycles of:
- amplifying said target sequence of nucleic acid by hybridization, replication; and
denaturing of nucleic acid, wherein denaturation is produced by applying a voltage to a solution containing said nucleic acid with an electrode so as to produce single-stranded nucleic acid in said solution not bound to said electrode. - View Dependent Claims (21, 27)
- amplifying said target sequence of nucleic acid by hybridization, replication; and
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22. A process for replicating a nucleic acid which comprises:
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separating strands of a sample double-stranded nucleic acid in solution under the influence of an electrical voltage applied to said solution from an electrode so as to produce single-stranded denatured nucleic acid in said solution not bound to said electrode; hybridizing said separated strands of said nucleic acid with at least one oligonucleotide primer that hybridizes with at least one of said strands of said denatured nucleic acid;
synthesizing at least one extension product of said at least one oligonucleotide primer which is sufficiently complementary to a respective strand of said nucleic acid to hybridize therewith; andseparating said at least one extension product from said nucleic acid strand with which it is hybridized to obtain said at least one extension product. - View Dependent Claims (23, 24, 25, 26)
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28. A process of detecting the presence or absence of a predetermined nucleic acid sequence in a sample which comprises:
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denaturing a sample double-stranded nucleic acid by means of a voltage applied to the sample in a solution by means of an electrode so as to produce single-stranded denatured nucleic acid in said solution not bound to said electrode; hybridizing said denatured nucleic acid with an oligonucleotide probe for said sequence; and determining whether the said hybridization has occurred.
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29. A kit for use in a process of nucleic acid amplification, said kit comprising:
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an electrode, a counter electrode, and more than one primer for use in a PCR procedure or an LCR procedure. - View Dependent Claims (30)
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Specification