Helicase-dependent amplification of RNA
First Claim
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1. A method for helicase-dependent amplification of an RNA, comprising:
- (a) adding to the RNA, a mixture comprising a reverse transcriptase, a helicase and a single strand binding protein unless the helicase is a thermostable helicase in which case the single strand binding protein is not required, a plurality of oligonucleotide primers and one or more polymerases; and
(b) in the mixture, reverse transcribing the RNA to form a cDNA and amplifying, by helicase-dependent amplification, the cDNA wherein the amplification does not occur in the absence of the helicase as determined by gel electrophoresis.
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Abstract
Methods and a kit are provided for selectively and exponentially amplifying nucleic acids and include the use of a single strand helicase preparation or a thermostable helicase in the absence of a single strand binding protein and a DNA polymerase such that the amplification can be performed isothermally.
89 Citations
7 Claims
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1. A method for helicase-dependent amplification of an RNA, comprising:
(a) adding to the RNA, a mixture comprising a reverse transcriptase, a helicase and a single strand binding protein unless the helicase is a thermostable helicase in which case the single strand binding protein is not required, a plurality of oligonucleotide primers and one or more polymerases; and
(b) in the mixture, reverse transcribing the RNA to form a cDNA and amplifying, by helicase-dependent amplification, the cDNA wherein the amplification does not occur in the absence of the helicase as determined by gel electrophoresis.- View Dependent Claims (2, 3, 4, 5, 6, 7)
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Current AssigneeBioHelix Corp. (Quidel Corporation)
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Original AssigneeNew England Biolabs Incorporated
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InventorsXu, Yan, Kong, Huimin, Vincent, Myriam
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Primary Examiner(s)Kim; Young J
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Assistant Examiner(s)Woolwine; Samuel C
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Application NumberUS11/327,175Publication NumberTime in Patent Office1,503 DaysField of SearchNoneUS Class Current435/91.2CPC Class CodesC12N 15/1096 cDNA Synthesis; Subtracted ...C12P 19/34 Polynucleotides, e.g. nucle...
