USRE39007E1 - Isothermal strand displacement nucleic acid amplification - Google Patents
Isothermal strand displacement nucleic acid amplification Download PDFInfo
- Publication number
- USRE39007E1 USRE39007E1 US10/164,833 US16483302A USRE39007E US RE39007 E1 USRE39007 E1 US RE39007E1 US 16483302 A US16483302 A US 16483302A US RE39007 E USRE39007 E US RE39007E
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- United States
- Prior art keywords
- nucleic acid
- complementary
- sequence
- primer
- primers
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Abstract
Description
-
- The Qβ replicase (QβR) method described in PCT Pub- lication No. 87-06270 and U.S. Pat. No. 4,786,600 uses a specific RNA probe which is capable of specific transcrip- tion by a replicase enzyme. The method requires the design and synthesis of RNA probes with replicase initiation sites.
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- The process includes the steps of priming single-stranded template nucleic acid strands with an excess of random oligonucleotide primers and incubating the single-stranded template nucleic acid strands and excess ran- dom oligonucleotide primers in the presence of excess amounts of an inducing agent, a strand displacement agent, and nucleoside triphosphate substrates to ran- domly amplify nucleic acid strands.
- SEQ. ID. NO. 1: CGCGTCTAGT CCTACTCAGG TGTTG
- SEQ. ID. NO. 2: CCCCAGATTC AGACTAGG
- SEQ. ID. NO. 3: CGGTCATTGA CTAGTATTTA GCCTT
- SEQ. ID. NO. 4: TTTCTTCATA GTACTGGTTC ACT
- SEQ. ID. NO. 5: ATTTCACTCC CTTAACAAGG GTT
- SEQ. ID. NO. 6: TTGTAAGCTA CAGGTTTCAG GT
- SEQ. ID. NO. 7: ACGGGCAATT GGTCTGCGAC C
Six of the primers were sense primers (with respect to the rRNA target). These primers had the following nucleic acid sequences (written 5′ to 3′): - SEQ. ID. NO. 8: AGCAAAGACC CGGAGGTCCG AAT
- SEQ. ID. NO. 9: GAATACATAG GTATGCAAAG CGA
- SEQ. ID. NO. 10: CCTGCCGAAC TGAAACATCT TAGT
- SEQ. ID. NO. 11: CAGAGGAAAA GAAATCGAAG AGAT
- SEQ. ID. NO. 12: TGTAGGATTG AGGATAAAGG AT
- SEQ. ID. NO. 13: GGACTCCTAG TTGAACACAT CTG- GAAAGA
TABLE 1 | |||
Chlamydia rRNA (Moles) | RLU (mean) | ||
4 × 10−18 | 307893 | ||
4 × 10−19 | 54771 | ||
4 × 10−20 | 3644 | ||
4 × 10−23 | 2354 | ||
0 | 2307 | ||
- SEQ. ID. NO. 14 CTATAGGGAG ACCACAACGG TTT
- SEQ. ID. NO. 15 CTAGAAATAA TTTTGTTTAA CTT
- SEQ. ID. NO. 16 AGAAGGAGAT ATACATATGG CT
- SEQ. ID. NO. 17 CATGACTGGT GGACAGCAAA TG
and four primers having the same sense as the RNA transcript, having the following nucleic acid sequences (written 5′ to 3′): - SEQ. ID. NO. 18 GAGCGAACGC AGTCAGGA
- SEQ. ID. NO. 19 GAGAAGTGGT CACGGAGGTA CGAGCGAACG CAGT
- SEQ. ID. NO. 20 GGAGATGGAA CGTACCATGT
- SEQ. ID. NO. 21 GGGAACTGAG CGGATTTACC GC
TABLE 2 | ||||
Probe | Sample | RLU (mean) | ||
SEQ. ID. 22 + | (4 × 10−19 mole) | 240474 | ||
SEQ. ID. 27 | ||||
SEQ. ID. 23 | (4 × 10−19 mole) | 3091 | ||
SEQ. ID. 23 | no target | 2720 | ||
TABLE 3 | ||||||
Primer (SEQ. | 24 | 24 | 24 | 25 | 25 | 25 |
ID. NO)1 | ||||||
Time | 5 min | 15 min | 30 min | 5 min | 15 min | 30 min |
% Product | 4.3 | 5.8 | 5.9 | 18 | 25 | 35 |
1In addition to the primers listed in Table 4, each reaction mixture contained a radiolabeled primer having the nucleotide sequence of SEQ. ID. NO. 19. |
Claims (42)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/164,833 USRE39007E1 (en) | 1994-03-16 | 2002-06-07 | Isothermal strand displacement nucleic acid amplification |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US21508194A | 1994-03-16 | 1994-03-16 | |
US08/972,799 US6087133A (en) | 1994-03-16 | 1997-11-18 | Isothermal strand displacement nucleic acid amplification |
US10/164,833 USRE39007E1 (en) | 1994-03-16 | 2002-06-07 | Isothermal strand displacement nucleic acid amplification |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/972,799 Reissue US6087133A (en) | 1994-03-16 | 1997-11-18 | Isothermal strand displacement nucleic acid amplification |
Publications (1)
Publication Number | Publication Date |
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USRE39007E1 true USRE39007E1 (en) | 2006-03-07 |
Family
ID=22801567
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/972,799 Ceased US6087133A (en) | 1994-03-16 | 1997-11-18 | Isothermal strand displacement nucleic acid amplification |
US09/506,282 Ceased US6214587B1 (en) | 1994-03-16 | 2000-02-17 | Isothermal strand displacement nucleic acid amplification |
US10/164,833 Expired - Lifetime USRE39007E1 (en) | 1994-03-16 | 2002-06-07 | Isothermal strand displacement nucleic acid amplification |
US10/165,281 Expired - Lifetime USRE38960E1 (en) | 1994-03-16 | 2002-06-07 | Isothermal strand displacement nucleic acid amplification |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/972,799 Ceased US6087133A (en) | 1994-03-16 | 1997-11-18 | Isothermal strand displacement nucleic acid amplification |
US09/506,282 Ceased US6214587B1 (en) | 1994-03-16 | 2000-02-17 | Isothermal strand displacement nucleic acid amplification |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/165,281 Expired - Lifetime USRE38960E1 (en) | 1994-03-16 | 2002-06-07 | Isothermal strand displacement nucleic acid amplification |
Country Status (11)
Country | Link |
---|---|
US (4) | US6087133A (en) |
EP (1) | EP0676476B1 (en) |
JP (1) | JPH09510351A (en) |
KR (1) | KR100230718B1 (en) |
AT (1) | ATE207543T1 (en) |
AU (1) | AU687535B2 (en) |
CA (1) | CA2185239C (en) |
DE (1) | DE69523360T2 (en) |
DK (1) | DK0676476T3 (en) |
ES (1) | ES2164112T3 (en) |
WO (1) | WO1995025180A1 (en) |
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Also Published As
Publication number | Publication date |
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AU2101695A (en) | 1995-10-03 |
US6087133A (en) | 2000-07-11 |
KR100230718B1 (en) | 1999-11-15 |
JPH09510351A (en) | 1997-10-21 |
EP0676476B1 (en) | 2001-10-24 |
CA2185239C (en) | 2002-12-17 |
DK0676476T3 (en) | 2001-12-03 |
KR970701791A (en) | 1997-04-12 |
US6214587B1 (en) | 2001-04-10 |
ATE207543T1 (en) | 2001-11-15 |
ES2164112T3 (en) | 2002-02-16 |
CA2185239A1 (en) | 1995-09-21 |
WO1995025180A1 (en) | 1995-09-21 |
EP0676476A1 (en) | 1995-10-11 |
USRE38960E1 (en) | 2006-01-31 |
DE69523360T2 (en) | 2002-07-11 |
AU687535B2 (en) | 1998-02-26 |
DE69523360D1 (en) | 2001-11-29 |
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