Process for controlling contamination of nucleic acid amplificat

Details Process for controlling contamination of nucleic acid amplificat Process for controlling contamination of nucleic acid amplificat

1989-06-01

1991-07-30

Wax, Robert A.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 91, 435200, 435227, C12Q 168, C12P 1934, C12N 922, C12N 978

Patent

active

050359967

ABSTRACT:
In the process according to this invention, an amplification procedure is performed on a first sample in which one or more of the four normal ribonucleoside triphosphates (rNTPs) or deoxyribonucleoside triphosphates (dNTPs) is replaced with an exo-sample nucleotide. After amplification, any contaminating amplified product that may be remaining is subjected to a physical, chemical, enzymatic, or biological treatment which renders nucleic acid containing the exo-sample nucleotide substantially unamplifiable. The treatment may be done as a separate step or it may be done in the presence of a second sample containing nucleic acid sequences to be amplified. The amplified nucleic acid sequences derived from the first sample which contaminate the second sample are not further substantially amplified during amplification of nucleic acid sequences of the second sample.

REFERENCES:
patent: 4683202 (1987-07-01), Mullis
Kwok and Higuchi, "Avoiding False Positives of PCR", Nature, vol. 339, (1989) pp. 237-238.
Saiki et al., "Primer-Directed Enzymatic . . . ", Science, vol. 239, (1988), 487-491.
Schaaper et al., PNAS, "Infidelity of DNA Synthesis Associated . . . ", vol. 80, pp. 487-491 (1983).

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