For increasing, quantitatively and the method for qualification nucleic acid

For increasing, quantitatively and the method for qualification nucleic acid

  • CN 101,278,061 B
  • Filed: 08/31/2006
  • Issued: 09/23/2015
  • Est. Priority Date: 09/01/2005
  • Status: Active Grant
First Claim
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1. increase the method for multiple selected nucleic acid molecule from nucleic acid molecule storehouse, and it comprises:

  • A () is increased multiple selected nucleic acid molecule in first round multiple reaction, described reaction comprises multiple external primers pair, often pair of selected nucleotide sequence of primer pair one has specificity and excessive existence in described reaction, wherein allows described reaction to proceed to reactive component, the point before significantly competition occur between amplicon;

    B () dilution enters from described first round multiple reaction described amplicon and the residual reaction components that multiple second takes turns amplified reaction, each described second takes turns amplified reaction all comprises at least one pair of internal primer, often pair of a kind of described selected nucleotide sequence of primer pair has specificity, wherein said dilution is 25 times of dilutions, the second further initiation taken turns in amplification is instructed by described internal primer substantially, and only increase described selected nucleotide sequence thus substantially that caused by described internal primer;

    WithC () takes turns in amplified reaction described multiple second the described selected nucleic acid molecule that increases further, make each second to take turns reaction by the subgroup in described the internal primer respectively described multiple selected nucleic acid molecule of further amplification.

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