JP7365363B2 - 方法 - Google Patents
方法 Download PDFInfo
- Publication number
- JP7365363B2 JP7365363B2 JP2020562111A JP2020562111A JP7365363B2 JP 7365363 B2 JP7365363 B2 JP 7365363B2 JP 2020562111 A JP2020562111 A JP 2020562111A JP 2020562111 A JP2020562111 A JP 2020562111A JP 7365363 B2 JP7365363 B2 JP 7365363B2
- Authority
- JP
- Japan
- Prior art keywords
- polynucleotide
- target
- adapter
- polynucleotides
- stranded
- 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.)
- Active
Links
- 238000000034 method Methods 0.000 title claims description 206
- 102000040430 polynucleotide Human genes 0.000 claims description 759
- 108091033319 polynucleotide Proteins 0.000 claims description 759
- 239000002157 polynucleotide Substances 0.000 claims description 759
- 108090000623 proteins and genes Proteins 0.000 claims description 219
- 102000004169 proteins and genes Human genes 0.000 claims description 186
- 239000012636 effector Substances 0.000 claims description 181
- 102000053602 DNA Human genes 0.000 claims description 179
- 108020004414 DNA Proteins 0.000 claims description 179
- 238000012163 sequencing technique Methods 0.000 claims description 152
- 238000003776 cleavage reaction Methods 0.000 claims description 126
- 230000007017 scission Effects 0.000 claims description 124
- 108091033409 CRISPR Proteins 0.000 claims description 119
- 239000002773 nucleotide Substances 0.000 claims description 91
- 125000003729 nucleotide group Chemical group 0.000 claims description 91
- 102000004190 Enzymes Human genes 0.000 claims description 61
- 108090000790 Enzymes Proteins 0.000 claims description 61
- 108010006785 Taq Polymerase Proteins 0.000 claims description 39
- 102000003960 Ligases Human genes 0.000 claims description 27
- 108090000364 Ligases Proteins 0.000 claims description 27
- 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 claims description 24
- 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 claims description 24
- 230000000694 effects Effects 0.000 claims description 14
- 230000003993 interaction Effects 0.000 claims description 12
- -1 Cas13 Proteins 0.000 claims description 9
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 claims description 9
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 claims description 9
- 238000012544 monitoring process Methods 0.000 claims description 9
- 108700004991 Cas12a Proteins 0.000 claims description 8
- 101150069031 CSN2 gene Proteins 0.000 claims description 5
- 101150074775 Csf1 gene Proteins 0.000 claims description 5
- 101150055601 cops2 gene Proteins 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 3
- 238000010168 coupling process Methods 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 2
- 239000000523 sample Substances 0.000 description 159
- 239000000203 mixture Substances 0.000 description 89
- 239000011324 bead Substances 0.000 description 82
- 210000004027 cell Anatomy 0.000 description 55
- 108010081734 Ribonucleoproteins Proteins 0.000 description 54
- 102000004389 Ribonucleoproteins Human genes 0.000 description 54
- 239000012634 fragment Substances 0.000 description 48
- 230000000295 complement effect Effects 0.000 description 47
- 238000005516 engineering process Methods 0.000 description 44
- 238000006243 chemical reaction Methods 0.000 description 42
- 239000000872 buffer Substances 0.000 description 41
- 241000588724 Escherichia coli Species 0.000 description 40
- 239000011148 porous material Substances 0.000 description 40
- 229920002477 rna polymer Polymers 0.000 description 36
- 238000003199 nucleic acid amplification method Methods 0.000 description 33
- 230000003321 amplification Effects 0.000 description 32
- 239000012528 membrane Substances 0.000 description 30
- 238000011534 incubation Methods 0.000 description 29
- 108020005004 Guide RNA Proteins 0.000 description 25
- 230000002779 inactivation Effects 0.000 description 25
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 22
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 22
- 101100310856 Drosophila melanogaster spri gene Proteins 0.000 description 22
- 230000030609 dephosphorylation Effects 0.000 description 22
- 238000006209 dephosphorylation reaction Methods 0.000 description 22
- 238000001847 surface plasmon resonance imaging Methods 0.000 description 22
- 108010017826 DNA Polymerase I Proteins 0.000 description 21
- 102000004594 DNA Polymerase I Human genes 0.000 description 21
- 108091028113 Trans-activating crRNA Proteins 0.000 description 21
- 102000039446 nucleic acids Human genes 0.000 description 20
- 108020004707 nucleic acids Proteins 0.000 description 20
- 238000012545 processing Methods 0.000 description 20
- 108060004795 Methyltransferase Proteins 0.000 description 19
- 108060002716 Exonuclease Proteins 0.000 description 18
- 102000013165 exonuclease Human genes 0.000 description 18
- 150000007523 nucleic acids Chemical class 0.000 description 18
- 238000002360 preparation method Methods 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 17
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 17
- 239000008188 pellet Substances 0.000 description 17
- 239000006228 supernatant Substances 0.000 description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
- 238000013459 approach Methods 0.000 description 16
- 244000309466 calf Species 0.000 description 16
- 230000000968 intestinal effect Effects 0.000 description 16
- 102000012410 DNA Ligases Human genes 0.000 description 15
- 108010061982 DNA Ligases Proteins 0.000 description 15
- 238000005259 measurement Methods 0.000 description 15
- 239000006148 magnetic separator Substances 0.000 description 14
- 108091034117 Oligonucleotide Proteins 0.000 description 13
- 238000000746 purification Methods 0.000 description 13
- 229910019142 PO4 Inorganic materials 0.000 description 12
- 230000003197 catalytic effect Effects 0.000 description 12
- 235000021317 phosphate Nutrition 0.000 description 12
- 201000010099 disease Diseases 0.000 description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 11
- 108010042407 Endonucleases Proteins 0.000 description 10
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 10
- 239000002342 ribonucleoside Substances 0.000 description 10
- 238000001712 DNA sequencing Methods 0.000 description 9
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 9
- 241000193996 Streptococcus pyogenes Species 0.000 description 9
- 238000010354 CRISPR gene editing Methods 0.000 description 8
- 238000010453 CRISPR/Cas method Methods 0.000 description 8
- 101710163270 Nuclease Proteins 0.000 description 8
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 8
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 8
- 238000001816 cooling Methods 0.000 description 8
- 239000005549 deoxyribonucleoside Substances 0.000 description 8
- 238000000605 extraction Methods 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 102100031780 Endonuclease Human genes 0.000 description 7
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 7
- 230000036425 denaturation Effects 0.000 description 7
- 238000004925 denaturation Methods 0.000 description 7
- 230000003544 deproteinization Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 230000035772 mutation Effects 0.000 description 7
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 230000007026 protein scission Effects 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 6
- 238000007672 fourth generation sequencing Methods 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 229920000642 polymer Polymers 0.000 description 6
- 229920001184 polypeptide Polymers 0.000 description 6
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 102000004196 processed proteins & peptides Human genes 0.000 description 6
- 230000002441 reversible effect Effects 0.000 description 6
- 125000006850 spacer group Chemical group 0.000 description 6
- 239000011534 wash buffer Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 5
- 108091093037 Peptide nucleic acid Proteins 0.000 description 5
- 101710183280 Topoisomerase Proteins 0.000 description 5
- 230000009471 action Effects 0.000 description 5
- 230000000692 anti-sense effect Effects 0.000 description 5
- 235000012000 cholesterol Nutrition 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 5
- 230000001404 mediated effect Effects 0.000 description 5
- 239000002777 nucleoside Substances 0.000 description 5
- 238000005580 one pot reaction Methods 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 5
- 239000010452 phosphate Substances 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 102000014914 Carrier Proteins Human genes 0.000 description 4
- 102000004533 Endonucleases Human genes 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 239000002202 Polyethylene glycol Substances 0.000 description 4
- 108020004682 Single-Stranded DNA Proteins 0.000 description 4
- 241000269841 Thunnus albacares Species 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 4
- 108091008324 binding proteins Proteins 0.000 description 4
- 239000012472 biological sample Substances 0.000 description 4
- 239000000356 contaminant Substances 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 239000012149 elution buffer Substances 0.000 description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 108091070501 miRNA Proteins 0.000 description 4
- 239000002679 microRNA Substances 0.000 description 4
- 150000003833 nucleoside derivatives Chemical class 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 229920001223 polyethylene glycol Polymers 0.000 description 4
- 230000000717 retained effect Effects 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- YZEUHQHUFTYLPH-UHFFFAOYSA-N 2-nitroimidazole Chemical compound [O-][N+](=O)C1=NC=CN1 YZEUHQHUFTYLPH-UHFFFAOYSA-N 0.000 description 3
- NEJMFSBXFBFELK-UHFFFAOYSA-N 4-nitro-1h-benzimidazole Chemical compound [O-][N+](=O)C1=CC=CC2=C1N=CN2 NEJMFSBXFBFELK-UHFFFAOYSA-N 0.000 description 3
- LAVZKLJDKGRZJG-UHFFFAOYSA-N 4-nitro-1h-indole Chemical compound [O-][N+](=O)C1=CC=CC2=C1C=CN2 LAVZKLJDKGRZJG-UHFFFAOYSA-N 0.000 description 3
- XORHNJQEWQGXCN-UHFFFAOYSA-N 4-nitro-1h-pyrazole Chemical compound [O-][N+](=O)C=1C=NNC=1 XORHNJQEWQGXCN-UHFFFAOYSA-N 0.000 description 3
- WSGURAYTCUVDQL-UHFFFAOYSA-N 5-nitro-1h-indazole Chemical compound [O-][N+](=O)C1=CC=C2NN=CC2=C1 WSGURAYTCUVDQL-UHFFFAOYSA-N 0.000 description 3
- OZFPSOBLQZPIAV-UHFFFAOYSA-N 5-nitro-1h-indole Chemical compound [O-][N+](=O)C1=CC=C2NC=CC2=C1 OZFPSOBLQZPIAV-UHFFFAOYSA-N 0.000 description 3
- PSWCIARYGITEOY-UHFFFAOYSA-N 6-nitro-1h-indole Chemical compound [O-][N+](=O)C1=CC=C2C=CNC2=C1 PSWCIARYGITEOY-UHFFFAOYSA-N 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 108700035208 EC 7.-.-.- Proteins 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 241001646716 Escherichia coli K-12 Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 101150043003 Htt gene Proteins 0.000 description 3
- 108010010677 Phosphodiesterase I Proteins 0.000 description 3
- 101800000684 Ribonuclease H Proteins 0.000 description 3
- 239000007984 Tris EDTA buffer Substances 0.000 description 3
- NLTUCYMLOPLUHL-KQYNXXCUSA-N adenosine 5'-[gamma-thio]triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=S)[C@@H](O)[C@H]1O NLTUCYMLOPLUHL-KQYNXXCUSA-N 0.000 description 3
- 230000004888 barrier function Effects 0.000 description 3
- 230000002457 bidirectional effect Effects 0.000 description 3
- 239000000090 biomarker Substances 0.000 description 3
- 230000002596 correlated effect Effects 0.000 description 3
- 230000000875 corresponding effect Effects 0.000 description 3
- 210000005260 human cell Anatomy 0.000 description 3
- 238000009396 hybridization Methods 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- 238000005304 joining Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 101150072531 10 gene Proteins 0.000 description 2
- NZJKEQFPRPAEPO-UHFFFAOYSA-N 1h-benzimidazol-4-amine Chemical compound NC1=CC=CC2=C1N=CN2 NZJKEQFPRPAEPO-UHFFFAOYSA-N 0.000 description 2
- VGONTNSXDCQUGY-RRKCRQDMSA-N 2'-deoxyinosine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC2=O)=C2N=C1 VGONTNSXDCQUGY-RRKCRQDMSA-N 0.000 description 2
- LOJNBPNACKZWAI-UHFFFAOYSA-N 3-nitro-1h-pyrrole Chemical compound [O-][N+](=O)C=1C=CNC=1 LOJNBPNACKZWAI-UHFFFAOYSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 2
- 241000057444 Lactobacillus brevis subsp. coagulans Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 108091078917 RecA family Proteins 0.000 description 2
- 102000041820 RecA family Human genes 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 102000008579 Transposases Human genes 0.000 description 2
- 108010020764 Transposases Proteins 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 150000001345 alkine derivatives Chemical class 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 150000001540 azides Chemical class 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 125000003636 chemical group Chemical group 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 230000009918 complex formation Effects 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 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 2
- 230000003247 decreasing effect Effects 0.000 description 2
- VGONTNSXDCQUGY-UHFFFAOYSA-N desoxyinosine Natural products C1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 VGONTNSXDCQUGY-UHFFFAOYSA-N 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 125000001924 fatty-acyl group Chemical group 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 229960003786 inosine Drugs 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 102000054765 polymorphisms of proteins Human genes 0.000 description 2
- 101150106848 rnp-2 gene Proteins 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- WETFHJRYOTYZFD-YIZRAAEISA-N (2r,3s,5s)-2-(hydroxymethyl)-5-(3-nitropyrrol-1-yl)oxolan-3-ol Chemical compound C1[C@H](O)[C@@H](CO)O[C@@H]1N1C=C([N+]([O-])=O)C=C1 WETFHJRYOTYZFD-YIZRAAEISA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 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 description 1
- XMTQQYYKAHVGBJ-UHFFFAOYSA-N 3-(3,4-DICHLOROPHENYL)-1,1-DIMETHYLUREA Chemical compound CN(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 XMTQQYYKAHVGBJ-UHFFFAOYSA-N 0.000 description 1
- LSMBOEFDMAIXTM-UUOKFMHZSA-N 7-[(2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-1h-imidazo[4,5-d]triazin-4-one Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NN=NC(O)=C2N=C1 LSMBOEFDMAIXTM-UUOKFMHZSA-N 0.000 description 1
- QFFLRMDXYQOYKO-KVQBGUIXSA-N 7-[(2r,4s,5r)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-1h-imidazo[4,5-d]triazin-4-one Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NN=NC(O)=C2N=C1 QFFLRMDXYQOYKO-KVQBGUIXSA-N 0.000 description 1
- 208000035657 Abasia Diseases 0.000 description 1
- 102100032187 Androgen receptor Human genes 0.000 description 1
- 241000203069 Archaea Species 0.000 description 1
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 244000075850 Avena orientalis Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 240000002791 Brassica napus Species 0.000 description 1
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 101150037468 CPD1 gene Proteins 0.000 description 1
- 238000010443 CRISPR/Cpf1 gene editing Methods 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 240000007154 Coffea arabica Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 108010043461 Deep Vent DNA polymerase Proteins 0.000 description 1
- 101100300807 Drosophila melanogaster spn-A gene Proteins 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 108010007577 Exodeoxyribonuclease I Proteins 0.000 description 1
- 102100029075 Exonuclease 1 Human genes 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 101150082209 Fmr1 gene Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000219146 Gossypium Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000775732 Homo sapiens Androgen receptor Proteins 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 238000006736 Huisgen cycloaddition reaction Methods 0.000 description 1
- 208000023105 Huntington disease Diseases 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 240000004322 Lens culinaris Species 0.000 description 1
- 235000014647 Lens culinaris subsp culinaris Nutrition 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 244000070406 Malus silvestris Species 0.000 description 1
- 108700011259 MicroRNAs Proteins 0.000 description 1
- 101100108853 Mus musculus Anp32e gene Proteins 0.000 description 1
- 240000005561 Musa balbisiana Species 0.000 description 1
- 206010068871 Myotonic dystrophy Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 101100221809 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cpd-7 gene Proteins 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 101100165815 Oryza sativa subsp. japonica CYP90A3 gene Proteins 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 102000009097 Phosphorylases Human genes 0.000 description 1
- 108010073135 Phosphorylases Proteins 0.000 description 1
- 241000276427 Poecilia reticulata Species 0.000 description 1
- 229920002594 Polyethylene Glycol 8000 Polymers 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 102000001218 Rec A Recombinases Human genes 0.000 description 1
- 108010055016 Rec A Recombinases Proteins 0.000 description 1
- 101100490727 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) AIF1 gene Proteins 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 108091081021 Sense strand Proteins 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 235000009470 Theobroma cacao Nutrition 0.000 description 1
- 244000299461 Theobroma cacao Species 0.000 description 1
- 101000865057 Thermococcus litoralis DNA polymerase Proteins 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 208000006269 X-Linked Bulbo-Spinal Atrophy Diseases 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 125000002015 acyclic group Chemical group 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 210000004381 amniotic fluid Anatomy 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 235000021016 apples Nutrition 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 125000004069 aziridinyl group Chemical group 0.000 description 1
- 235000021015 bananas Nutrition 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 125000000640 cyclooctyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C([H])([H])C1([H])[H] 0.000 description 1
- 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 1
- 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 1
- 230000006378 damage Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 101150025236 dmaW gene Proteins 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 150000002118 epoxides Chemical class 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 108010052305 exodeoxyribonuclease III Proteins 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 238000003505 heat denaturation Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000002847 impedance measurement Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 239000012038 nucleophile Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 125000000714 pyrimidinyl group Chemical group 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 101150059622 rnp3 gene Proteins 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000005783 single-strand break Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000000707 stereoselective effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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
- C12Q1/6869—Methods for sequencing
-
- 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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB1808554.8A GB201808554D0 (en) | 2018-05-24 | 2018-05-24 | Method |
| GB1808554.8 | 2018-05-24 | ||
| PCT/GB2019/051444 WO2019224560A1 (en) | 2018-05-24 | 2019-05-24 | Method |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2021523704A JP2021523704A (ja) | 2021-09-09 |
| JP2021523704A5 JP2021523704A5 (https=) | 2022-04-25 |
| JP7365363B2 true JP7365363B2 (ja) | 2023-10-19 |
Family
ID=62812420
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2020562111A Active JP7365363B2 (ja) | 2018-05-24 | 2019-05-24 | 方法 |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US20210198732A1 (https=) |
| EP (2) | EP4549584A3 (https=) |
| JP (1) | JP7365363B2 (https=) |
| CN (1) | CN112105744A (https=) |
| AU (1) | AU2019274949B2 (https=) |
| CA (1) | CA3096856A1 (https=) |
| GB (1) | GB201808554D0 (https=) |
| WO (1) | WO2019224560A1 (https=) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11046995B2 (en) | 2016-08-16 | 2021-06-29 | The Regents Of The University Of California | Method for finding low abundance sequences by hybridization (FLASH) |
| EP3731959B1 (en) | 2017-12-29 | 2025-10-08 | Clear Labs, Inc. | Automated priming and library loading device |
| TW202039845A (zh) * | 2018-12-20 | 2020-11-01 | 北京大學 | 使用加標籤的嚮導rna構建體進行高效基因篩選的組合物和方法 |
| CN112029838B (zh) * | 2020-07-23 | 2022-07-12 | 东南大学 | 一种用于DNA均相检测的CRISPR/Cas9分型PCR方法及其应用 |
| LT4211260T (lt) | 2020-09-11 | 2025-06-25 | New England Biolabs, Inc. | Imobilizuotų fermentų taikymas nanoporų bibliotekos konstravimui |
| WO2022243437A1 (en) * | 2021-05-19 | 2022-11-24 | KWS SAAT SE & Co. KGaA | Sample preparation with oppositely oriented guide polynucleotides |
| WO2023107899A2 (en) * | 2021-12-07 | 2023-06-15 | Caribou Biosciences, Inc. | A method of capturing crispr endonuclease cleavage products |
| CN114457145B (zh) * | 2022-01-29 | 2023-08-11 | 成都齐碳科技有限公司 | 用于表征靶多核苷酸测序的接头、构建体、方法和应用 |
| CN114921533A (zh) * | 2022-05-05 | 2022-08-19 | 成都齐碳科技有限公司 | 用于表征目标多核苷酸的方法及衔接体 |
| CN114921535B (zh) * | 2022-05-19 | 2023-05-23 | 四川大学华西医院 | 一种rna长片段靶向测序方法 |
| CN114921534B (zh) * | 2022-05-19 | 2023-05-30 | 四川大学华西医院 | 一种高靶向效率和数据产量的rna长片段靶向测序方法 |
| WO2024138517A1 (zh) * | 2022-12-29 | 2024-07-04 | 深圳华大生命科学研究院 | 提升测序通量的文库接头设计 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017044843A1 (en) | 2015-09-11 | 2017-03-16 | The General Hospital Corporation | Full interrogation of nuclease dsbs and sequencing (find-seq) |
Family Cites Families (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6267872B1 (en) | 1998-11-06 | 2001-07-31 | The Regents Of The University Of California | Miniature support for thin films containing single channels or nanopores and methods for using same |
| FR2861405A1 (fr) * | 2003-10-24 | 2005-04-29 | Centre Nat Rech Scient | METHODE DE PRODUCTION D'ADNc PLEINE LONGUEUR PAR LIGATURATION D'ADAPTATEUR A EXTREMITE COHESIVE |
| WO2005124888A1 (en) | 2004-06-08 | 2005-12-29 | President And Fellows Of Harvard College | Suspended carbon nanotube field effect transistor |
| US20110121840A1 (en) | 2007-02-20 | 2011-05-26 | Gurdial Singh Sanghera | Lipid Bilayer Sensor System |
| KR20110125226A (ko) | 2009-01-30 | 2011-11-18 | 옥스포드 나노포어 테크놀로지즈 리미티드 | 혼성화 링커 |
| EP4737389A2 (en) | 2011-05-27 | 2026-05-06 | Oxford Nanopore Technologies PLC | Coupling method |
| CN104039979B (zh) | 2011-10-21 | 2016-08-24 | 牛津纳米孔技术公司 | 使用孔和Hel308解旋酶表征目标多核苷酸的酶方法 |
| US9617591B2 (en) | 2011-12-29 | 2017-04-11 | Oxford Nanopore Technologies Ltd. | Method for characterising a polynucleotide by using a XPD helicase |
| AU2012360244B2 (en) | 2011-12-29 | 2018-08-23 | Oxford Nanopore Technologies Limited | Enzyme method |
| JP6429773B2 (ja) | 2012-07-19 | 2018-11-28 | オックスフォード ナノポール テクノロジーズ リミテッド | 酵素構築物 |
| CA2879261C (en) | 2012-07-19 | 2022-12-06 | Oxford Nanopore Technologies Limited | Modified helicases |
| US11155860B2 (en) | 2012-07-19 | 2021-10-26 | Oxford Nanopore Technologies Ltd. | SSB method |
| CA2901545C (en) | 2013-03-08 | 2019-10-08 | Oxford Nanopore Technologies Limited | Use of spacer elements in a nucleic acid to control movement of a helicase |
| CN117947149A (zh) | 2013-10-18 | 2024-04-30 | 牛津纳米孔科技公开有限公司 | 经修饰的酶 |
| WO2015150786A1 (en) | 2014-04-04 | 2015-10-08 | Oxford Nanopore Technologies Limited | Method for characterising a double stranded nucleic acid using a nano-pore and anchor molecules at both ends of said nucleic acid |
| GB201417712D0 (en) | 2014-10-07 | 2014-11-19 | Oxford Nanopore Tech Ltd | Method |
| EP3183358B1 (en) * | 2014-08-19 | 2020-10-07 | President and Fellows of Harvard College | Rna-guided systems for probing and mapping of nucleic acids |
| GB201418469D0 (en) | 2014-10-17 | 2014-12-03 | Oxford Nanopore Tech Ltd | Method |
| WO2017162754A1 (en) * | 2016-03-22 | 2017-09-28 | Vib Vzw | Means and methods for amplifying nucleotide sequences |
| US11046995B2 (en) * | 2016-08-16 | 2021-06-29 | The Regents Of The University Of California | Method for finding low abundance sequences by hybridization (FLASH) |
| GB201616590D0 (en) * | 2016-09-29 | 2016-11-16 | Oxford Nanopore Technologies Limited | Method |
-
2018
- 2018-05-24 GB GBGB1808554.8A patent/GB201808554D0/en not_active Ceased
-
2019
- 2019-05-24 US US17/057,863 patent/US20210198732A1/en active Pending
- 2019-05-24 CA CA3096856A patent/CA3096856A1/en active Pending
- 2019-05-24 WO PCT/GB2019/051444 patent/WO2019224560A1/en not_active Ceased
- 2019-05-24 EP EP25161085.3A patent/EP4549584A3/en active Pending
- 2019-05-24 AU AU2019274949A patent/AU2019274949B2/en active Active
- 2019-05-24 CN CN201980029513.6A patent/CN112105744A/zh active Pending
- 2019-05-24 EP EP19727476.4A patent/EP3802862A1/en not_active Ceased
- 2019-05-24 JP JP2020562111A patent/JP7365363B2/ja active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017044843A1 (en) | 2015-09-11 | 2017-03-16 | The General Hospital Corporation | Full interrogation of nuclease dsbs and sequencing (find-seq) |
Non-Patent Citations (1)
| Title |
|---|
| MASAYUKI FUKASAWA; ET AL，MICROARRAY ANALYSIS OF PROMOTER METHYLATION IN LUNG CANCERS，JOURNAL OF HUMAN GENETICS，TO，SPRINGER-VERLAG，2006年04月，VOL:51, NR:4，PAGE(S):368 - 374，http://dx.doi.org/10.1007/s10038-005-0355-4 |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2019224560A1 (en) | 2019-11-28 |
| CN112105744A (zh) | 2020-12-18 |
| AU2019274949B2 (en) | 2025-08-14 |
| EP3802862A1 (en) | 2021-04-14 |
| US20210198732A1 (en) | 2021-07-01 |
| EP4549584A2 (en) | 2025-05-07 |
| JP2021523704A (ja) | 2021-09-09 |
| AU2019274949A1 (en) | 2020-10-15 |
| CA3096856A1 (en) | 2019-11-28 |
| GB201808554D0 (en) | 2018-07-11 |
| EP4549584A3 (en) | 2025-07-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7365363B2 (ja) | 方法 | |
| US20250109426A1 (en) | Compositions and methods for targeted depletion, enrichment, and partitioning of nucleic acids using crispr/cas system proteins | |
| US11459602B2 (en) | Method for generating double stranded DNA libraries and sequencing methods for the identification of methylated cytosines | |
| EP2510126B1 (en) | Multi-sample indexing for multiplex genotyping | |
| EP3386550B1 (en) | Methods for the making and using of guide nucleic acids | |
| EP3635114B1 (en) | Creation and use of guide nucleic acids | |
| CN104080958A (zh) | 用于定向核酸扩增和测序的组合物和方法 | |
| JP7766029B2 (ja) | 共有結合で閉端された核酸分子末端を使用したｎｇｓライブラリー調製 | |
| US20220403368A1 (en) | Methods and systems for preparing a nucleic acid construct for single molecule characterisation | |
| EP3768855B1 (en) | Methods and compositions for recombinase-mediated selective cleavage of nucleic acids | |
| JP2002537774A (ja) | 多型ｄｎａフラグメントおよびその使用 | |
| WO2024209000A1 (en) | Linkers for duplex sequencing | |
| US20250137041A1 (en) | Means and method for fluorescent in situ hybridization using crispr-cas9 variants and a nuclease | |
| HK40038012B (en) | Methods and compositions for recombinase-mediated selective cleavage of nucleic acids | |
| HK40038012A (en) | Methods and compositions for recombinase-mediated selective cleavage of nucleic acids | |
| HK1176096B (en) | Multi-sample indexing for multiplex genotyping | |
| HK1176096A (en) | Multi-sample indexing for multiplex genotyping |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220414 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20220414 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230418 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230714 |
|
| 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: 20230912 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20231006 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7365363 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |