JP5020074B2 - 新規ホットスタート核酸増幅法 - Google Patents
新規ホットスタート核酸増幅法 Download PDFInfo
- Publication number
- JP5020074B2 JP5020074B2 JP2007519541A JP2007519541A JP5020074B2 JP 5020074 B2 JP5020074 B2 JP 5020074B2 JP 2007519541 A JP2007519541 A JP 2007519541A JP 2007519541 A JP2007519541 A JP 2007519541A JP 5020074 B2 JP5020074 B2 JP 5020074B2
- Authority
- JP
- Japan
- Prior art keywords
- temperature
- nucleic acid
- reaction
- primer
- reaction mixture
- Prior art date
- 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 - Fee Related
Links
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 27
- 238000006243 chemical reaction Methods 0.000 claims abstract description 198
- 101710088729 Single-stranded nucleic acid-binding protein Proteins 0.000 claims abstract description 121
- 238000000034 method Methods 0.000 claims abstract description 96
- 239000000203 mixture Substances 0.000 claims abstract description 78
- 239000011541 reaction mixture Substances 0.000 claims abstract description 68
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 64
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 56
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 56
- 230000003321 amplification Effects 0.000 claims abstract description 26
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 4
- 238000009396 hybridization Methods 0.000 claims description 42
- 108090000623 proteins and genes Proteins 0.000 claims description 34
- 230000000295 complement effect Effects 0.000 claims description 29
- 102000004169 proteins and genes Human genes 0.000 claims description 27
- 230000027455 binding Effects 0.000 claims description 25
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 21
- 102000004190 Enzymes Human genes 0.000 claims description 19
- 108090000790 Enzymes Proteins 0.000 claims description 19
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 238000004925 denaturation Methods 0.000 claims description 12
- 230000036425 denaturation Effects 0.000 claims description 12
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 11
- 230000002401 inhibitory effect Effects 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 11
- 230000008569 process Effects 0.000 claims description 11
- 239000002773 nucleotide Substances 0.000 claims description 9
- 125000003729 nucleotide group Chemical group 0.000 claims description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 8
- 150000001413 amino acids Chemical class 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 238000001668 nucleic acid synthesis Methods 0.000 claims description 7
- 150000001768 cations Chemical class 0.000 claims description 3
- 230000003362 replicative effect Effects 0.000 claims description 3
- 230000001404 mediated effect Effects 0.000 claims 5
- 239000000047 product Substances 0.000 description 82
- 108020004414 DNA Proteins 0.000 description 76
- 108010006785 Taq Polymerase Proteins 0.000 description 52
- 101710126859 Single-stranded DNA-binding protein Proteins 0.000 description 42
- 101710093976 Plasmid-derived single-stranded DNA-binding protein Proteins 0.000 description 38
- 235000019527 sweetened beverage Nutrition 0.000 description 38
- 102000053602 DNA Human genes 0.000 description 34
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 31
- 230000000694 effects Effects 0.000 description 27
- 239000000872 buffer Substances 0.000 description 25
- 239000012536 storage buffer Substances 0.000 description 18
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 15
- 238000011534 incubation Methods 0.000 description 15
- 238000003752 polymerase chain reaction Methods 0.000 description 15
- 239000011780 sodium chloride Substances 0.000 description 15
- 230000037452 priming Effects 0.000 description 14
- 241000588724 Escherichia coli Species 0.000 description 12
- 230000003993 interaction Effects 0.000 description 12
- 230000002441 reversible effect Effects 0.000 description 10
- 229940088598 enzyme Drugs 0.000 description 9
- 239000000178 monomer Substances 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 230000006820 DNA synthesis Effects 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 239000000539 dimer Substances 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 238000002844 melting Methods 0.000 description 8
- 230000008018 melting Effects 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000007795 chemical reaction product Substances 0.000 description 7
- 108020004682 Single-Stranded DNA Proteins 0.000 description 6
- 230000001351 cycling effect Effects 0.000 description 6
- 238000003757 reverse transcription PCR Methods 0.000 description 6
- 238000012408 PCR amplification Methods 0.000 description 5
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 5
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 5
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 5
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 5
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 102000014914 Carrier Proteins Human genes 0.000 description 4
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 4
- 101710176276 SSB protein Proteins 0.000 description 4
- 238000000137 annealing Methods 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 108091008324 binding proteins Proteins 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 4
- 239000005547 deoxyribonucleotide Substances 0.000 description 4
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 238000007849 hot-start PCR Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 238000013207 serial dilution Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 3
- 102000052510 DNA-Binding Proteins Human genes 0.000 description 3
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000701832 Enterobacteria phage T3 Species 0.000 description 3
- 102100034343 Integrase Human genes 0.000 description 3
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- 239000008049 TAE buffer Substances 0.000 description 3
- HGEVZDLYZYVYHD-UHFFFAOYSA-N acetic acid;2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound CC(O)=O.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O HGEVZDLYZYVYHD-UHFFFAOYSA-N 0.000 description 3
- 239000011543 agarose gel Substances 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 229960000723 ampicillin Drugs 0.000 description 3
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000011109 contamination Methods 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 3
- 229960005542 ethidium bromide Drugs 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 238000010839 reverse transcription Methods 0.000 description 3
- 238000012552 review Methods 0.000 description 3
- 230000009919 sequestration Effects 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 2
- 108700020911 DNA-Binding Proteins Proteins 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 2
- -1 Mg 2+ Chemical class 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- 229920000669 heparin Polymers 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000011901 isothermal amplification Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000011533 pre-incubation Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000011535 reaction buffer Substances 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 108010005008 single stranded DNA dependent ATPase Proteins 0.000 description 2
- 108700014590 single-stranded DNA binding proteins Proteins 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- 101000852101 Bombyx mori DNA topoisomerase 2 Proteins 0.000 description 1
- FRPHFZCDPYBUAU-UHFFFAOYSA-N Bromocresolgreen Chemical compound CC1=C(Br)C(O)=C(Br)C=C1C1(C=2C(=C(Br)C(O)=C(Br)C=2)C)C2=CC=CC=C2S(=O)(=O)O1 FRPHFZCDPYBUAU-UHFFFAOYSA-N 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 101710116602 DNA-Binding protein G5P Proteins 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 101000740205 Homo sapiens Sal-like protein 1 Proteins 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 101150008132 NDE1 gene Proteins 0.000 description 1
- 206010029719 Nonspecific reaction Diseases 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108010002747 Pfu DNA polymerase Proteins 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 102000001218 Rec A Recombinases Human genes 0.000 description 1
- 108010055016 Rec A Recombinases Proteins 0.000 description 1
- 101710162453 Replication factor A Proteins 0.000 description 1
- 101710176758 Replication protein A 70 kDa DNA-binding subunit Proteins 0.000 description 1
- 102100022380 SOSS complex subunit B2 Human genes 0.000 description 1
- 102100037204 Sal-like protein 1 Human genes 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000589596 Thermus Species 0.000 description 1
- 108030003004 Triphosphatases Proteins 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000027832 depurination Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 108700025694 p53 Genes Proteins 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 108010083127 phage repressor proteins Proteins 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
Images
Classifications
-
- 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
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
-
- 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
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Peptides Or Proteins (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US58436204P | 2004-06-30 | 2004-06-30 | |
| US60/584,362 | 2004-06-30 | ||
| PCT/US2005/023824 WO2006005074A2 (en) | 2004-06-30 | 2005-06-30 | Novel hot start nucleic acid amplification |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2008516584A JP2008516584A (ja) | 2008-05-22 |
| JP2008516584A5 JP2008516584A5 (https=) | 2009-07-23 |
| JP5020074B2 true JP5020074B2 (ja) | 2012-09-05 |
Family
ID=35783411
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007519541A Expired - Fee Related JP5020074B2 (ja) | 2004-06-30 | 2005-06-30 | 新規ホットスタート核酸増幅法 |
Country Status (7)
| Country | Link |
|---|---|
| US (3) | US7700281B2 (https=) |
| EP (2) | EP2366802A1 (https=) |
| JP (1) | JP5020074B2 (https=) |
| AT (1) | ATE527380T1 (https=) |
| CA (1) | CA2569394A1 (https=) |
| NO (1) | NO20065470L (https=) |
| WO (1) | WO2006005074A2 (https=) |
Families Citing this family (56)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7700281B2 (en) * | 2004-06-30 | 2010-04-20 | Usb Corporation | Hot start nucleic acid amplification |
| AU2007268075C1 (en) | 2006-06-01 | 2013-03-07 | Trilink Biotechnologies | Chemically modified oligonucleotide primers for nucleic acid amplification |
| JP2010537637A (ja) | 2007-08-27 | 2010-12-09 | アプライド バイオシステムズ インコーポレイテッド | Ｐｃｒのための方法および組成物 |
| JP5843614B2 (ja) | 2009-01-30 | 2016-01-13 | オックスフォード ナノポア テクノロジーズ リミテッド | 膜貫通配列決定における核酸構築物のためのアダプター |
| JP5687414B2 (ja) * | 2009-04-20 | 2015-03-18 | オリンパス株式会社 | 多型の識別方法 |
| KR101162422B1 (ko) * | 2009-11-30 | 2012-07-10 | 박수민 | 핵산 증폭용 조성물 |
| US9597188B2 (en) | 2011-04-06 | 2017-03-21 | DePuy Synthes Products, Inc. | Version-replicating instrument and orthopaedic surgical procedure for using the same to implant a revision hip prosthesis |
| AU2012288629B2 (en) | 2011-07-25 | 2017-02-02 | Oxford Nanopore Technologies Limited | Hairpin loop method for double strand polynucleotide sequencing using transmembrane pores |
| CN114214394A (zh) * | 2012-06-08 | 2022-03-22 | 爱奥尼安技术公司 | 核苷酸扩增反应 |
| KR20210072137A (ko) | 2012-06-14 | 2021-06-16 | 라이프 테크놀로지스 코포레이션 | 폴리머라제 연쇄 반응 (pcr)을 위한 신규 조성물, 방법 및 키트 |
| US11155860B2 (en) * | 2012-07-19 | 2021-10-26 | Oxford Nanopore Technologies Ltd. | SSB method |
| US10273541B2 (en) | 2012-08-14 | 2019-04-30 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US10752949B2 (en) | 2012-08-14 | 2020-08-25 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| CA3216609C (en) | 2012-08-14 | 2024-05-14 | 10X Genomics, Inc. | Microcapsule compositions and methods |
| US10221442B2 (en) | 2012-08-14 | 2019-03-05 | 10X Genomics, Inc. | Compositions and methods for sample processing |
| US9701998B2 (en) | 2012-12-14 | 2017-07-11 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US11591637B2 (en) | 2012-08-14 | 2023-02-28 | 10X Genomics, Inc. | Compositions and methods for sample processing |
| US9951386B2 (en) | 2014-06-26 | 2018-04-24 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US10323279B2 (en) | 2012-08-14 | 2019-06-18 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US10584381B2 (en) | 2012-08-14 | 2020-03-10 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| WO2014093676A1 (en) | 2012-12-14 | 2014-06-19 | 10X Technologies, Inc. | Methods and systems for processing polynucleotides |
| US10533221B2 (en) | 2012-12-14 | 2020-01-14 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| AU2014214682B2 (en) | 2013-02-08 | 2018-07-26 | 10X Genomics, Inc. | Polynucleotide barcode generation |
| GB201314695D0 (en) | 2013-08-16 | 2013-10-02 | Oxford Nanopore Tech Ltd | Method |
| KR102168813B1 (ko) | 2013-03-08 | 2020-10-22 | 옥스포드 나노포어 테크놀로지즈 리미티드 | 효소 정지 방법 |
| GB201403096D0 (en) | 2014-02-21 | 2014-04-09 | Oxford Nanopore Tech Ltd | Sample preparation method |
| WO2015157567A1 (en) | 2014-04-10 | 2015-10-15 | 10X Genomics, Inc. | Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same |
| US12312640B2 (en) | 2014-06-26 | 2025-05-27 | 10X Genomics, Inc. | Analysis of nucleic acid sequences |
| MX2016016904A (es) | 2014-06-26 | 2017-03-27 | 10X Genomics Inc | Analisis de secuencias de acidos nucleicos. |
| CN113249435B (zh) | 2014-06-26 | 2024-09-03 | 10X基因组学有限公司 | 分析来自单个细胞或细胞群体的核酸的方法 |
| MX2017005267A (es) | 2014-10-29 | 2017-07-26 | 10X Genomics Inc | Metodos y composiciones para la secuenciacion de acidos nucleicos seleccionados como diana. |
| US9975122B2 (en) | 2014-11-05 | 2018-05-22 | 10X Genomics, Inc. | Instrument systems for integrated sample processing |
| SG11201705615UA (en) | 2015-01-12 | 2017-08-30 | 10X Genomics Inc | Processes and systems for preparing nucleic acid sequencing libraries and libraries prepared using same |
| EP4286516A3 (en) | 2015-02-24 | 2024-03-06 | 10X Genomics, Inc. | Partition processing methods and systems |
| BR112017018054A2 (pt) | 2015-02-24 | 2018-07-24 | 10X Genomics Inc | métodos para a cobertura de sequências de ácidos nucleicos direcionadas |
| SG11201804086VA (en) | 2015-12-04 | 2018-06-28 | 10X Genomics Inc | Methods and compositions for nucleic acid analysis |
| US10626383B2 (en) | 2016-01-15 | 2020-04-21 | Thermo Fisher Scientific Baltics Uab | Thermophilic DNA polymerase mutants |
| WO2017197343A2 (en) | 2016-05-12 | 2017-11-16 | 10X Genomics, Inc. | Microfluidic on-chip filters |
| WO2017197338A1 (en) | 2016-05-13 | 2017-11-16 | 10X Genomics, Inc. | Microfluidic systems and methods of use |
| GB201609220D0 (en) | 2016-05-25 | 2016-07-06 | Oxford Nanopore Tech Ltd | Method |
| US10815525B2 (en) | 2016-12-22 | 2020-10-27 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US10011872B1 (en) | 2016-12-22 | 2018-07-03 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| US10550429B2 (en) | 2016-12-22 | 2020-02-04 | 10X Genomics, Inc. | Methods and systems for processing polynucleotides |
| CN110214186B (zh) | 2017-01-30 | 2023-11-24 | 10X基因组学有限公司 | 用于基于微滴的单细胞条形编码的方法和系统 |
| US12264411B2 (en) | 2017-01-30 | 2025-04-01 | 10X Genomics, Inc. | Methods and systems for analysis |
| CN109526228B (zh) | 2017-05-26 | 2022-11-25 | 10X基因组学有限公司 | 转座酶可接近性染色质的单细胞分析 |
| US10844372B2 (en) | 2017-05-26 | 2020-11-24 | 10X Genomics, Inc. | Single cell analysis of transposase accessible chromatin |
| JP7111706B2 (ja) | 2017-06-07 | 2022-08-02 | タカラバイオ株式会社 | オリゴヌクレオチドの保存方法 |
| US11618891B2 (en) | 2017-06-26 | 2023-04-04 | Thermo Fisher Scientific Baltics Uab | Thermophilic DNA polymerase mutants |
| EP3954782A1 (en) | 2017-11-15 | 2022-02-16 | 10X Genomics, Inc. | Functionalized gel beads |
| US10829815B2 (en) | 2017-11-17 | 2020-11-10 | 10X Genomics, Inc. | Methods and systems for associating physical and genetic properties of biological particles |
| CN112262218B (zh) | 2018-04-06 | 2024-11-08 | 10X基因组学有限公司 | 用于单细胞处理中的质量控制的系统和方法 |
| GB201807793D0 (en) | 2018-05-14 | 2018-06-27 | Oxford Nanopore Tech Ltd | Method |
| KR102529958B1 (ko) * | 2021-01-18 | 2023-05-08 | 가톨릭대학교 산학협력단 | 저 gc 함량 dna의 정량적 증폭을 위한 pcr 첨가제 조성물 |
| CN116004776A (zh) * | 2022-08-30 | 2023-04-25 | 上海伯杰医疗科技股份有限公司 | 一管式RT-qPCR全预混反应试剂、应用及RT-qPCR方法 |
| WO2025071292A1 (ko) * | 2023-09-26 | 2025-04-03 | 주식회사 씨젠 | 핫 스타트 핵산 중합효소의 성능을 평가하는 방법 |
Family Cites Families (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61502282A (ja) | 1984-06-01 | 1986-10-09 | ナショナル・バイオメディカル・リサ−チ・ファウンデ−ション | 酵素剤を用いた触媒作用による核酸交雑方法 |
| US4683195A (en) | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
| US4965188A (en) | 1986-08-22 | 1990-10-23 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences using a thermostable enzyme |
| US4889818A (en) | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
| US4994372A (en) | 1987-01-14 | 1991-02-19 | President And Fellows Of Harvard College | DNA sequencing |
| US5646019A (en) * | 1989-10-24 | 1997-07-08 | Stratagene | Method for producing primed nucleic acid templates |
| US5605824A (en) * | 1989-10-24 | 1997-02-25 | Stratagene | Composition for hybridizing nucleic acids using single-stranded nucleic acid binding protein |
| WO1991006679A1 (en) | 1989-10-24 | 1991-05-16 | Stratagene | An improved method for hybridizing nucleic acids using single-stranded nucleic acid binding protein |
| US5449603A (en) * | 1989-10-24 | 1995-09-12 | Stratagene | Method for hybridizing nucleic acids using single-stranded nucleic acid binding protein |
| DE69131891T2 (de) * | 1990-02-16 | 2000-06-15 | F. Hoffmann-La Roche Ag, Basel | Verbesserungen in der spezifität und zweckmässigkeit der polymerase-kettenreaktion |
| US5223414A (en) * | 1990-05-07 | 1993-06-29 | Sri International | Process for nucleic acid hybridization and amplification |
| AU645915B2 (en) * | 1991-07-23 | 1994-01-27 | F. Hoffmann-La Roche Ag | Improvements in the in situ PCR |
| WO1993015226A1 (en) * | 1992-01-28 | 1993-08-05 | President And Fellows Of Harvard College | Method for nucleic acid hybridization using single-stranded dna binding protein |
| US5338671A (en) * | 1992-10-07 | 1994-08-16 | Eastman Kodak Company | DNA amplification with thermostable DNA polymerase and polymerase inhibiting antibody |
| US5364790A (en) * | 1993-02-16 | 1994-11-15 | The Perkin-Elmer Corporation | In situ PCR amplification system |
| US5773258A (en) * | 1995-08-25 | 1998-06-30 | Roche Molecular Systems, Inc. | Nucleic acid amplification using a reversibly inactivated thermostable enzyme |
| US5854033A (en) * | 1995-11-21 | 1998-12-29 | Yale University | Rolling circle replication reporter systems |
| EP0869187A3 (en) | 1997-02-24 | 2001-02-28 | Becton, Dickinson and Company | Replication of nucleic acids using single-strand DNA binding proteins |
| US6124120A (en) * | 1997-10-08 | 2000-09-26 | Yale University | Multiple displacement amplification |
| US6183998B1 (en) * | 1998-05-29 | 2001-02-06 | Qiagen Gmbh Max-Volmer-Strasse 4 | Method for reversible modification of thermostable enzymes |
| GB9920194D0 (en) * | 1999-08-27 | 1999-10-27 | Advanced Biotech Ltd | A heat-stable thermostable DNA polymerase for use in nucleic acid amplification |
| WO2003008624A2 (en) * | 2001-07-15 | 2003-01-30 | Keck Graduate Institute | Nucleic acid amplification using nicking agents |
| US7399590B2 (en) * | 2002-02-21 | 2008-07-15 | Asm Scientific, Inc. | Recombinase polymerase amplification |
| WO2004022770A2 (en) | 2002-09-05 | 2004-03-18 | Invitrogen Corporation | Compositions and methods for synthesizing nucleic acids |
| JP4632788B2 (ja) * | 2002-09-20 | 2011-02-16 | ニュー・イングランド・バイオラブズ・インコーポレイティッド | 核酸のヘリカーゼ依存性増幅 |
| JP2004113108A (ja) * | 2002-09-26 | 2004-04-15 | Aisin Seiki Co Ltd | Ｄｎａ回収法 |
| JP2005110621A (ja) * | 2003-10-10 | 2005-04-28 | Aisin Seiki Co Ltd | 核酸増幅方法及び核酸増幅用試薬キット |
| US7759057B2 (en) * | 2004-02-04 | 2010-07-20 | Ut-Battelle, Llc | Method for analyzing microbial communities |
| US7700281B2 (en) * | 2004-06-30 | 2010-04-20 | Usb Corporation | Hot start nucleic acid amplification |
-
2005
- 2005-06-29 US US11/171,008 patent/US7700281B2/en active Active
- 2005-06-30 CA CA002569394A patent/CA2569394A1/en not_active Abandoned
- 2005-06-30 EP EP11163362A patent/EP2366802A1/en not_active Withdrawn
- 2005-06-30 WO PCT/US2005/023824 patent/WO2006005074A2/en not_active Ceased
- 2005-06-30 EP EP05764400A patent/EP1778866B1/en not_active Expired - Lifetime
- 2005-06-30 AT AT05764400T patent/ATE527380T1/de not_active IP Right Cessation
- 2005-06-30 JP JP2007519541A patent/JP5020074B2/ja not_active Expired - Fee Related
-
2006
- 2006-11-28 NO NO20065470A patent/NO20065470L/no not_active Application Discontinuation
-
2007
- 2007-11-14 US US11/939,820 patent/US7951534B2/en not_active Expired - Lifetime
-
2011
- 2011-03-29 US US13/074,752 patent/US8404443B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JP2008516584A (ja) | 2008-05-22 |
| NO20065470L (no) | 2007-01-26 |
| ATE527380T1 (de) | 2011-10-15 |
| EP2366802A1 (en) | 2011-09-21 |
| EP1778866B1 (en) | 2011-10-05 |
| CA2569394A1 (en) | 2006-01-12 |
| WO2006005074A2 (en) | 2006-01-12 |
| US20060029952A1 (en) | 2006-02-09 |
| US7700281B2 (en) | 2010-04-20 |
| EP1778866A4 (en) | 2009-06-03 |
| EP1778866A2 (en) | 2007-05-02 |
| US8404443B2 (en) | 2013-03-26 |
| US20110189736A1 (en) | 2011-08-04 |
| US20080138878A1 (en) | 2008-06-12 |
| WO2006005074A3 (en) | 2008-06-19 |
| US7951534B2 (en) | 2011-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5020074B2 (ja) | 新規ホットスタート核酸増幅法 | |
| US5512462A (en) | Methods and reagents for the polymerase chain reaction amplification of long DNA sequences | |
| US5773257A (en) | Method for producing primed nucleic acid templates | |
| JP3867926B2 (ja) | 核酸の増幅法 | |
| US20010049125A1 (en) | End-complementary polymerase reaction | |
| JP2002291490A (ja) | 修飾プライマーを使用する増幅 | |
| WO1994003624A1 (en) | Methods for the isothermal amplification of nucleic acid molecules | |
| JPH10513058A (ja) | 核酸分子の等温増幅方法 | |
| US20080044921A1 (en) | Primers used in novel gene amplification method | |
| US5605824A (en) | Composition for hybridizing nucleic acids using single-stranded nucleic acid binding protein | |
| JP4718493B2 (ja) | 核酸増幅中のプライマー凝集体形成を減少させるためのｄＵＴＰに基づく組成物 | |
| WO1991006679A1 (en) | An improved method for hybridizing nucleic acids using single-stranded nucleic acid binding protein | |
| US20110111462A1 (en) | Composition and Method for Synthesizing a Deoxyribonucleotide Chain Using a Double Stranded Nucleic Acid Complex with a Thermostable Polymerase | |
| WO2002090538A1 (en) | Method of synthesizing nucleic acid | |
| JP4942160B2 (ja) | ＲｅｃＡタンパク質を利用した核酸の等温増幅法 | |
| JP2005318884A (ja) | 核酸の増幅方法 | |
| JPH06253843A (ja) | Ｄｎａ配列の増幅法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20080627 |
|
| A524 | Written submission of copy of amendment under article 19 pct |
Free format text: JAPANESE INTERMEDIATE CODE: A524 Effective date: 20090602 |
|
| A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20100915 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110308 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20110608 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20110615 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110708 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20120117 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120417 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20120515 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20120612 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 5020074 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150622 Year of fee payment: 3 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| LAPS | Cancellation because of no payment of annual fees |