EP0505012A2
Expired / Lapsed
EP
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Means for amplifying nucleic acid sequences
Assignee: F Hoffmann La Roche AG
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Filing: 1986-03-27
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Granted: —
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Expires: 2006-03-27
Abstract
Abstract The present invention is directed to a means for exponentially amplifying at least one specific nucleic acid sequence contained in a nucleic acid or a mixture of nucleic acids wherein each nucleic acid consists of two separate complementary strands, of equal or unequal length, or is single-stranded, which means comprise primer for each different specific nucleic acid sequence being amplified selected so as to provide a primer which is substantially complementary to each of the different strands of each specific sequence being amplified.
Inventors
- 👤 Kary Banks Mullis
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 22:20
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; B; B01; B01L; B01L7/00; B01L7/52; C07; C07K; C07K14/00; C07K14/795; C07K14/805; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6827; C12Q1/6844; C12Q1/6853; C12Q1/6858; C12Q1/686; B01J; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00495; B01J2219/00583; B01J2219/0059; B01J2219/00718; B01J2219/0072; B01J2219/00722; C12Q1/6876; C12Q1/6883; C12Q1/6886; C12Q1/6888; C12Q1/689; C40; C40B; C40B40/00; C40B40/04; C40B40/06; C40B60/00; C40B60/14
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
76 words
Means for exponentially amplifying at least one specific nucleic acid sequence contained in a nucleic acid or a mixture of nucleic acids wherein each nucleic acid consists of two separate complementary strands, of equal or unequal length, or is single-stranded, which means comprise primer for each different specific nucleic acid sequence being amplified selected so as to provide a primer which is substantially complementary to each of the different strands of each specific sequence being amplified.
Claim 2
Dependent Claim
19 words
Means according to claim 1 characterized in that said specific nucleic acid sequence is contained within a larger sequence.
Claim 3
Dependent Claim
36 words
Means according of claim 1 or 2 characterized in that said specific nucleic acid sequence is DNA or RNA, including messenger RNA, which DNA or RNA may be single-stranded or double-stranded, or is a DNA-RNA hybrid.
Claim 4
Dependent Claim
21 words
Means according to any one of claims 1 - 3 characterized in that said specific nucleic acid sequence is genomic DNA.
Claim 5
Dependent Claim
39 words
Means according to any one of claims 1 - 4 characterized by comprising a collection of primers for each strand of the specific nucleic acid being amplified, at least one of said primers is substantially complementary to said strand.
Claim 6
Dependent Claim
21 words
Means according to any one of claims 1 - 5 characterized by primer containing about 7 - 25 nucleotides or more.
Claim 7
Dependent Claim
40 words
Means according to any one of claims 1 - 6 characterized in that at least one primer contains a nucleotide sequence attached to the 5'-end which sequence is non-complementary to the corresponding strand of the specific nucleic acid being amplified.
Claim 8
Dependent Claim
43 words
Means according to any one of claims 1 - 6 characterized in that at least one primer is interspersed with bases or a nucleotide sequence which bases or nucleotide sequence are non-complementary to the corresponding strand of the specific nucleic acid being amplified.
Claim 9
Dependent Claim
23 words
Means according to claim 7 or 8 characterized in that the non-complementary nucleotide sequence is a promoter, a linker or a coding sequence.
Claim 10
Dependent Claim
28 words
Means according to any one of claims 1 - 9 characterized by further comprising an inducing agent which will function to accomplish the synthesis of primer extension products.
Claim 11
Dependent Claim
44 words
Means according to claim 10 characterized in that the inducing agent is an enzyme selected from E. coli DNA polymerase I, Klenow fragment of E. coli DNA polymerase I, T4 DNA polymerase, another DNA polymerase, a reverse transcriptase and other enzymes including heat-stable enzymes.
Claim 12
Dependent Claim
16 words
Means according to any one of claims 1 - 11 characterized by further comprising different nucleotides.
Claim 13
Dependent Claim
11 words
Means according to claim 12 characterized by comprising readily detectable nucleotides.
Claim 14
Dependent Claim
22 words
Means according to any one of claims 1 - 13 characterized by further comprising an enzyme from the class known as helicases.
Claim 15
Dependent Claim
52 words
The use of the means according to any one of claims 1 - 14 for amplifying at least one specific nucleic acid sequence contained in a nucleic acid or a mixture of nucleic acids wherein each nucleic acid consists of two separate complementary strands, of equal or unequal length, or is single-stranded.
Claim 16
Dependent Claim
35 words
The use of the means according to any one of claims 1 - 14 for improving the efficiency of cloning a particular nucleic acid such as DNA or messenger RNA into a suitable expression vector.
Claim 17
Dependent Claim
23 words
The use of the means according to any one of claims 1 - 14 for amplifying a target sequence to facilitate detection thereof.
Claim 18
Dependent Claim
34 words
The use of the means according to any one of claims 1 - 14 for obtaining large amounts of the desired sequence from a mixture of nucleic acids resulting from an imperfect chemical synthesis.
Claim 19
Dependent Claim
23 words
The use of the means according to any one of claims 1 - 14 for introducing in vitro mutations into a specific sequence.
Claim 20
Dependent Claim
58 words
The use of the means according to any one of claims 1 - 14 for enabling detection and/or characterization of specific nucleic acid sequences associated with infectious diseases such as those caused by bacteria, viruses and protozoan parasites, genetic disorders such as those caused by specific deletions and/or mutations in genomic DNA or cellular disorders such as cancer.
References Cited (Backward Citations)
No citation records indexed.
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