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Method of minimizing contamination in amplification reactions using a reaction tube with a penetrable membrane

  • US 5,604,101 A
  • Filed: 11/17/1995
  • Issued: 02/18/1997
  • Est. Priority Date: 10/22/1993
  • Status: Expired due to Term
First Claim
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1. In a method for amplifying and detecting nucleic acid materials comprising the steps of:

  • a) adding a sample suspected to contain a target nucleic acid material to an amplification vessel along with oligonucleotide probes or primers, at least one of which bears a detectable reporter group, for amplification of said suspected target nucleic acid to form a reaction mixture;

    b) sealing the reaction mixture inside said vessel by closing a tightly sealing cap;

    c) amplifying the target nucleic acid material within said vessel; and

    d) detecting the presence of amplified target nucleic acid by detection of said detectable reporter group;

    wherein the improvement comprisesi) providing said sealing cap with a membrane that is penetrable by a pipettor probe;

    ii) removing a portion of the reaction mixture from said vessel for detection wherein said removing is effected by piercing said cap membrane with a pipettor probe aspirating said portion of the reaction mixture into said pipettor; and

    iii) dispensing said portion in a distinct detection compartment without uncapping said vessel, thereby avoiding drops or aerosols of the amplified material which might contaminate the environment, unreacted samples or reagents.

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