US5035996A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for controlling contamination of nucleic acid amplification reactions
Assignee: Life Technologies Corp
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Filing: 1989-06-01
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Granted: 1991-07-30
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Expires: 2009-06-01
Abstract
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.
Inventors
- 👤 James L. Hartley
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 17:26
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/708; C12Q1/68; C12Q1/6844; C12Q1/6848
Claims Analysis
Total Claims: 5
Claim 1
Independent Claim
93 words
A method for controlling contamination in sequential nucleic acid amplification processes comprising first and second nucleic acid amplification processes to amplify nucleic acid sequences in a first and a second sample, respectively, which comprises: carrying out said first nucleic acid amplification process on said nucleic acid sequence in said first sample in the presence of deoxyuridine triphosphate, to thereby generate a deoxyuracil-containing product of amplification; and treating said deoxyuracil-containing product of amplification with uracil DNA glycosylase prior to carrying out said second amplification process on said nucleic acid sequence in said second sample.
Claim 2
Independent Claim
93 words
A method for controlling contamination in sequential nucleic acid amplification processes comprising first and second nucleic acid amplification processes to amplify nucleic acid sequences in a first and a second sample, respectively, which comprises: carrying out said first nucleic acid amplification process on said nucleic acid sequence in said first sample in the presence of deoxyuridine triphosphate, to thereby generate a deoxyuracil-containing product of amplification; and treating said deoxyuracil-containing product of amplification with uracil DNA glycosylase prior to carrying out said second amplification process on said nucleic acid sequence in said second sample.
Claim 3
Dependent Claim
254 words
The process of claim 1 wherein said nucleic acid sequences in said first and second samples have two separate complementary strands each, wherein said first nucleic acid amplification process on said first sample comprises: (1) treating said strands with two oligonucleotide primers, for each different specific sequence being amplified, under conditions such that for each different sequence being amplified, an extension product of each primer is synthesized which is complementary to each nucleic acid strand and which incorporates deoxyuridine, wherein said primers are selected so as to be sufficiently complementary to different strands of each specific sequence to hybridize therewith, such that the extension product synthesized from one primer, when it is separated from its complement, can serve as a template for synthesis of the extension product of the other primer; (2) separating the primer extension products from the templates on which they are synthesized to produce single-stranded molecules; (3) treating said single-stranded molecules generated from step (2) with the primers of step (1) under conditions that a primer extension product is synthesized using each of the single strands produced in step (2) as a template; (4) repeating steps (1) through (3) at least once, thereby amplifying said specific nucleic acid sequence contained in said first sample; and wherein said second nucleic acid amplification process on said second sample comprises (5) treating said second sample with uracil DNA glycosylase; and (6) repeating steps (1) through (3) on said second sample at least once, thereby amplifying the nucleic acid sequence contained in said second sample.
Claim 4
Dependent Claim
40 words
The process of claim 1 wherein, after treating said deoxyuracil-containing product with uracil DNA glycosylase, and prior to carrying out said second amplification process, the second sample is heated so as to terminate the action of said uracil DNA glycosylase.
Claim 5
Dependent Claim
28 words
The process of claim 2 wherein, after said step (5), the process comprises: (7) heating said sample so as to terminate the action of said uracil DNA glycosylase.
References Cited (Backward Citations)
US4683202A
Cited
US4683202B1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-07-29 | 1995-01-28 | 1995-07-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-29 | 1999-01-28 | 1999-07-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-29 | 2003-01-28 | 2003-07-30 | $7700.00 | Window Closed / Lapsed |