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Method for amplification and detection of RNA and DNA sequences

  • US 5,783,391 A
  • Filed: 07/12/1995
  • Issued: 07/21/1998
  • Est. Priority Date: 03/10/1988
  • Status: Expired due to Fees
First Claim
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1. The method which comprises:

  • (i) providing an RNA sample which may have a target nucleotide sequence;

    (ii) providing a first primer and a second primer for reverse transcriptase-polymerase chain reaction amplification of said RNA sampleat least one of said first and second primers having a 5'"'"' terminal moiety which is a recognition sequence for an RNA polymerase,(iii) subjecting said RNA sample to reverse transcriptase polymerase chain reaction amplification including thermal denaturation between cycles denaturation between cycles, to produce a first amplification product;

    said amplification product having said recognition sequence for an RNA polymerase of said at least one of said first and second primers,(iv) providing an RNA polymerase which recognizes the said recognition sequence of said at least one of said primers;

    (v) subjecting said first amplification product of step (iii) to transcription by said RNA polymerase of step (iv) to produce a second amplification product;

    (vi) subjecting said second amplification product of step (v) to reverse transcriptase polymerase chain reaction amplification to produce a third amplification product;

    (vii) determining whether said target nucleic acid sequence is present in said third amplification product.

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