US5043272A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Amplification of nucleic acid sequences using oligonucleotides of random sequence as primers
Assignee: Life Technologies Inc
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Filing: 1989-04-27
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Granted: 1991-08-27
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Expires: 2009-04-27
Abstract
Abstract According to this invention, a process for substantially amplifying template nucleic acid present in a sample is described, wherein said amplification may be performed without prior knowledge of specific sequences, which process comprises apposition of random oligonucleotide primers to said template nucleic acid under conditions such that extension products of said primers are synthesized which are complementary to said template nucleic acid.
Inventors
- 👤 James L. Hartley
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 06:24
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/708; C12Q1/68; C12Q1/6844; C12Q1/6858; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
77 words
A process for substantially amplifying template nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising the steps of: (a) priming template nucleic acid strands with an excess of random oligonucleotide primers; and (b) incubating said template nucleic acid strands and said excess random oligonucleotide primers in the presence of an excess of an inducing agent, a strand displacement agent, and an excess of triphosphate substrates to randomly amplify nucleic acid strands.
Claim 2
Dependent Claim
77 words
A process for substantially amplifying template nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising the steps of: (a) priming template nucleic acid strands with an excess of random oligonucleotide primers; and (b) incubating said template nucleic acid strands and said excess random oligonucleotide primers in the presence of an excess of an inducing agent, a strand displacement agent, and an excess of triphosphate substrates to randomly amplify nucleic acid strands.
Claim 3
Dependent Claim
15 words
The process of claim 1, wherein said oligonucleotide primer is less than 50 bases long.
Claim 4
Dependent Claim
13 words
The process of claim 2, wherein said oligonucleotide primer is 4-20 bases long.
Claim 5
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 10 bases long.
Claim 6
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 9 bases long.
Claim 7
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 8 bases long.
Claim 8
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 7 bases long.
Claim 9
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 6 bases long.
Claim 10
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 5 bases long.
Claim 11
Dependent Claim
13 words
The process of claim 3, wherein said oligonucleotide primer is 4 bases long.
Claim 12
Dependent Claim
20 words
The process of claim 1, wherein said inducing agent and said strand displacement agent are activities of the same protein.
Claim 13
Dependent Claim
13 words
The process of claim 1, wherein said process is catalyzed by an enzyme.
Claim 14
Dependent Claim
38 words
The process of claim 12, wherein said enzyme is selected from the group consisting of E. coli DNA polymerase I, the Klenow fragment of E. coli DNA polymerase I, bacteriophage T5 DNA polymerase, and bacteriophage T7 DNA polymerase.
Claim 15
Dependent Claim
18 words
The process of claim 13, wherein said enzyme is the Klenow fragment of E. coli DNA polymerase I.
Claim 16
Dependent Claim
14 words
The process of claim 1, wherein said sample is derived from a biological source.
Claim 17
Dependent Claim
20 words
The process of claim 15, wherein said biological source is derived from a human, animal, plant, bacterial or viral source.
Claim 18
Dependent Claim
14 words
The process of claim 16, wherein said viral source is human papilloma virus DNA.
Claim 19
Dependent Claim
12 words
The process of claim 16, wherein said viral source is HIV DNA.
Claim 20
Dependent Claim
20 words
The process of claim 15, wherein said sample is derived from a tissue, fluid or secretion of said biological source.
Claim 21
Dependent Claim
35 words
The process of claim 19, wherein said tissue, fluid or secretion is selected from the group consisting of blood, stool, sera, urine, saliva, tears, biopsy tissue sample, histology tissue sample, PAP smear, mole, and wart.
Claim 22
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 10-fold.
Claim 23
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 30-fold.
Claim 24
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 100-fold.
Claim 25
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 300-fold.
Claim 26
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 500-fold.
Claim 27
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 1000-fold.
Claim 28
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 5,000-fold.
Claim 29
Dependent Claim
12 words
The process of claim 1, wherein said amplification is at least 8,000-fold.
Claim 30
Dependent Claim
87 words
A process for substantially amplifying template nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising the steps of: (a) priming template nucleic acid strands with an excess of random oligonucleotide primers wherein said primers consist of 6-mers to 10 mers; and (b) incubating said template nucleic acid strands and said excess random oligonucleotide primers in the presence of an excess of the Klenow fragment of DNA Polymerase I and an excess of triphosphate substrates to randomly amplify template nucleic acid strands.
Claim 31
Dependent Claim
87 words
A process for substantially amplifying template nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising the steps of: (a) priming template nucleic acid strands with an excess of random oligonucleotide primers wherein said primers consist of 6-mers to 10 mers; and (b) incubating said template nucleic acid strands and said excess random oligonucleotide primers in the presence of an excess of the Klenow fragment of DNA Polymerase I and an excess of triphosphate substrates to randomly amplify template nucleic acid strands.
Claim 32
Dependent Claim
14 words
The process of claim 29, wherein said sample is derived from a biological source.
Claim 33
Dependent Claim
20 words
The process of claim 30, wherein said biological source is derived from a human, animal, plant, bacterial or viral source.
Claim 34
Dependent Claim
14 words
The process of claim 31, wherein said viral source is human papilloma virus DNA.
Claim 35
Dependent Claim
12 words
The process of claim 31, wherein said viral source is HIV DNA.
Claim 36
Dependent Claim
20 words
The process of claim 30, wherein said sample is derived from a tissue, fluid or secretion of said biological source.
Claim 37
Dependent Claim
35 words
The process of claim 34, wherein said tissue, fluid or secretion is selected from the group consisting of blood, stool, sera, urine, saliva, tears, biopsy tissue sample, histology tissue sample, PAP smear, mole, and wart.
Claim 38
Dependent Claim
46 words
A process for detecting a papilloma virus in a sample comprising: (a) substantial amplification of said papilloma virus nucleic acid sequences in said sample wherein said substantial amplification comprises a randomly primed but template dependent synthesis of papilloma virus DNA; and (b) detecting said papilloma virus.
Claim 39
Dependent Claim
46 words
A process for detecting a papilloma virus in a sample comprising: (a) substantial amplification of said papilloma virus nucleic acid sequences in said sample wherein said substantial amplification comprises a randomly primed but template dependent synthesis of papilloma virus DNA; and (b) detecting said papilloma virus.
Claim 40
Dependent Claim
21 words
The process of claim 36, wherein said randomly primed amplification occurs with a primer that is less than 50 bases long.
Claim 41
Dependent Claim
19 words
The process of claim 37, wherein said randomly primed amplification occurs with a primer that is 4-20 bases long.
Claim 42
Dependent Claim
19 words
The process of claim 38, wherein said randomly primed amplification occurs with a primer that is 8 bases long.
Claim 43
Dependent Claim
14 words
The process of claim 36, wherein said sample is derived from a biological source.
Claim 44
Dependent Claim
20 words
The process of claim 40, wherein said biological source is derived from a human, animal, plant, bacterial or viral source.
Claim 45
Dependent Claim
14 words
The process of claim 41, wherein said viral source is human papilloma virus DNA.
Claim 46
Dependent Claim
20 words
The process of claim 40, wherein said sample is derived from a tissue, fluid or secretion of said biological source.
Claim 47
Dependent Claim
35 words
The process of claim 43, wherein said tissue, fluid or secretion is selected from the group consisting of blood, stool, sera, urine, saliva, tears, biopsy tissue sample, histology tissue sample, PAP smear, mole, and wart.
Claim 48
Dependent Claim
85 words
A kit for substantially amplifying nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising a carrier being compartmentalized to receive in close confinement therein one or more containers wherein: (a) a first container or series of containers contains random oligonucleotide primers; (b) a second container contains an inducing agent; (c) a third container or series of containers contains triphosphate substrates; and (d) a fourth container or series of containers contains buffer for reconstituting or diluting components of said kit.
Claim 49
Dependent Claim
85 words
A kit for substantially amplifying nucleic acid sequences in a sample in a randomly primed and template dependent manner, comprising a carrier being compartmentalized to receive in close confinement therein one or more containers wherein: (a) a first container or series of containers contains random oligonucleotide primers; (b) a second container contains an inducing agent; (c) a third container or series of containers contains triphosphate substrates; and (d) a fourth container or series of containers contains buffer for reconstituting or diluting components of said kit.
Claim 50
Dependent Claim
14 words
The kit of claim 45, further comprising a container containing a strand displacement agent.
Claim 51
Dependent Claim
20 words
The kit of claim 45, further comprising a container containing a probe capable of identifying a bacterial or viral species.
Claim 52
Dependent Claim
14 words
The kit of claim 47, wherein said probe is a human papilloma virus probe.
Claim 53
Dependent Claim
12 words
The kit of claim 47, wherein said probe is a HIV probe.
Claim 54
Dependent Claim
12 words
The kit of claim 47, wherein said probe is a Salmonella probe.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-08-26 | 1995-02-25 | 1995-08-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-08-26 | 1999-02-25 | 1999-08-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-08-26 | 2003-02-25 | 2003-08-27 | $7700.00 | Window Closed / Lapsed |