JP2020524488A - 配列に基づく遺伝子試験のための対照を作製するための組成物及び方法 - Google Patents
配列に基づく遺伝子試験のための対照を作製するための組成物及び方法 Download PDFInfo
- Publication number
- JP2020524488A JP2020524488A JP2019566955A JP2019566955A JP2020524488A JP 2020524488 A JP2020524488 A JP 2020524488A JP 2019566955 A JP2019566955 A JP 2019566955A JP 2019566955 A JP2019566955 A JP 2019566955A JP 2020524488 A JP2020524488 A JP 2020524488A
- Authority
- JP
- Japan
- Prior art keywords
- adapter
- template
- nucleic acid
- universal
- amplification
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 292
- 239000000203 mixture Substances 0.000 title claims abstract description 89
- 238000012360 testing method Methods 0.000 title claims abstract description 75
- 230000002068 genetic effect Effects 0.000 title claims description 34
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 334
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 320
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 320
- 238000012163 sequencing technique Methods 0.000 claims abstract description 95
- 238000010200 validation analysis Methods 0.000 claims abstract description 21
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 230000003321 amplification Effects 0.000 claims description 215
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 215
- 125000003729 nucleotide group Chemical group 0.000 claims description 147
- 239000002773 nucleotide Substances 0.000 claims description 141
- 108020004414 DNA Proteins 0.000 claims description 82
- 239000012082 adaptor molecule Substances 0.000 claims description 72
- 230000000295 complement effect Effects 0.000 claims description 59
- 230000027455 binding Effects 0.000 claims description 57
- 238000006243 chemical reaction Methods 0.000 claims description 54
- 206010028980 Neoplasm Diseases 0.000 claims description 50
- 238000003752 polymerase chain reaction Methods 0.000 claims description 49
- 238000001514 detection method Methods 0.000 claims description 47
- 238000003776 cleavage reaction Methods 0.000 claims description 42
- 239000011324 bead Substances 0.000 claims description 41
- 108091093088 Amplicon Proteins 0.000 claims description 39
- 108091008146 restriction endonucleases Proteins 0.000 claims description 39
- 230000007017 scission Effects 0.000 claims description 39
- 230000008569 process Effects 0.000 claims description 33
- 208000036878 aneuploidy Diseases 0.000 claims description 25
- 231100001075 aneuploidy Toxicity 0.000 claims description 25
- 208000037280 Trisomy Diseases 0.000 claims description 24
- 239000003795 chemical substances by application Substances 0.000 claims description 23
- 239000012530 fluid Substances 0.000 claims description 23
- 239000013060 biological fluid Substances 0.000 claims description 20
- 238000011282 treatment Methods 0.000 claims description 20
- 210000004369 blood Anatomy 0.000 claims description 16
- 239000008280 blood Substances 0.000 claims description 16
- 210000003754 fetus Anatomy 0.000 claims description 15
- 201000010374 Down Syndrome Diseases 0.000 claims description 14
- 244000052769 pathogen Species 0.000 claims description 14
- 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 claims description 12
- 206010044688 Trisomy 21 Diseases 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 12
- 238000002405 diagnostic procedure Methods 0.000 claims description 12
- 230000002255 enzymatic effect Effects 0.000 claims description 12
- 239000003446 ligand Substances 0.000 claims description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 10
- 230000001605 fetal effect Effects 0.000 claims description 10
- 230000008774 maternal effect Effects 0.000 claims description 10
- 230000001717 pathogenic effect Effects 0.000 claims description 10
- 210000002700 urine Anatomy 0.000 claims description 10
- 206010036790 Productive cough Diseases 0.000 claims description 9
- 210000003802 sputum Anatomy 0.000 claims description 9
- 208000024794 sputum Diseases 0.000 claims description 9
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 8
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 8
- 208000035269 cancer or benign tumor Diseases 0.000 claims description 7
- 201000009928 Patau syndrome Diseases 0.000 claims description 6
- 206010044686 Trisomy 13 Diseases 0.000 claims description 6
- 208000006284 Trisomy 13 Syndrome Diseases 0.000 claims description 6
- 229960002685 biotin Drugs 0.000 claims description 6
- 235000020958 biotin Nutrition 0.000 claims description 6
- 239000011616 biotin Substances 0.000 claims description 6
- 230000001225 therapeutic effect Effects 0.000 claims description 5
- 206010053884 trisomy 18 Diseases 0.000 claims description 5
- 206010071547 Trisomy 9 Diseases 0.000 claims description 4
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 208000015181 infectious disease Diseases 0.000 claims description 4
- 238000010208 microarray analysis Methods 0.000 claims description 4
- 238000011269 treatment regimen Methods 0.000 claims description 4
- 206010044689 trisomy 22 Diseases 0.000 claims description 4
- 108020000946 Bacterial DNA Proteins 0.000 claims description 3
- 201000006360 Edwards syndrome Diseases 0.000 claims description 3
- 208000007159 Trisomy 18 Syndrome Diseases 0.000 claims description 3
- 206010053871 Trisomy 8 Diseases 0.000 claims description 3
- 108020005202 Viral DNA Proteins 0.000 claims description 3
- 208000034298 trisomy chromosome 8 Diseases 0.000 claims description 3
- 230000009452 underexpressoin Effects 0.000 claims description 3
- 244000052613 viral pathogen Species 0.000 claims description 3
- 244000052616 bacterial pathogen Species 0.000 claims description 2
- 239000013641 positive control Substances 0.000 abstract description 13
- 239000013642 negative control Substances 0.000 abstract description 11
- 238000004164 analytical calibration Methods 0.000 abstract 1
- 239000013615 primer Substances 0.000 description 207
- 239000000523 sample Substances 0.000 description 111
- 102000053602 DNA Human genes 0.000 description 78
- 239000012634 fragment Substances 0.000 description 54
- 239000002585 base Substances 0.000 description 50
- 210000000349 chromosome Anatomy 0.000 description 47
- 102000040430 polynucleotide Human genes 0.000 description 43
- 108091033319 polynucleotide Proteins 0.000 description 43
- 239000002157 polynucleotide Substances 0.000 description 43
- 210000002381 plasma Anatomy 0.000 description 34
- 230000002441 reversible effect Effects 0.000 description 34
- 239000000758 substrate Substances 0.000 description 29
- 239000003153 chemical reaction reagent Substances 0.000 description 27
- 239000000463 material Substances 0.000 description 26
- 238000012217 deletion Methods 0.000 description 24
- 230000037430 deletion Effects 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 23
- 239000007787 solid Substances 0.000 description 22
- 108090000623 proteins and genes Proteins 0.000 description 21
- 239000000047 product Substances 0.000 description 20
- 230000015572 biosynthetic process Effects 0.000 description 19
- 239000000126 substance Substances 0.000 description 19
- 108091028664 Ribonucleotide Proteins 0.000 description 17
- 238000000137 annealing Methods 0.000 description 17
- 230000004048 modification Effects 0.000 description 17
- 238000012986 modification Methods 0.000 description 17
- 239000002336 ribonucleotide Substances 0.000 description 17
- 125000002652 ribonucleotide group Chemical group 0.000 description 17
- 201000011510 cancer Diseases 0.000 description 16
- 239000000499 gel Substances 0.000 description 16
- 239000007790 solid phase Substances 0.000 description 16
- 230000005856 abnormality Effects 0.000 description 15
- 238000010348 incorporation Methods 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 208000035657 Abasia Diseases 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- -1 Mg ++ or Mn ++ Chemical class 0.000 description 12
- 108091034117 Oligonucleotide Proteins 0.000 description 12
- 239000012472 biological sample Substances 0.000 description 12
- 108010042407 Endonucleases Proteins 0.000 description 11
- 238000005516 engineering process Methods 0.000 description 11
- 230000007717 exclusion Effects 0.000 description 11
- 102000004169 proteins and genes Human genes 0.000 description 11
- 239000002096 quantum dot Substances 0.000 description 11
- 229920002477 rna polymer Polymers 0.000 description 11
- 208000031639 Chromosome Deletion Diseases 0.000 description 10
- 239000000872 buffer Substances 0.000 description 10
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 10
- 230000029087 digestion Effects 0.000 description 10
- 239000000178 monomer Substances 0.000 description 10
- 208000030454 monosomy Diseases 0.000 description 10
- 102100031780 Endonuclease Human genes 0.000 description 9
- 230000008901 benefit Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 238000013467 fragmentation Methods 0.000 description 9
- 238000006062 fragmentation reaction Methods 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 8
- 208000026310 Breast neoplasm Diseases 0.000 description 8
- 108010090804 Streptavidin Proteins 0.000 description 8
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 8
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 8
- 238000003556 assay Methods 0.000 description 8
- 239000005547 deoxyribonucleotide Substances 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- 210000003296 saliva Anatomy 0.000 description 8
- 208000011580 syndromic disease Diseases 0.000 description 8
- IOSROLCFSUFOFE-UHFFFAOYSA-L 2-nitro-1h-imidazole;platinum(2+);dichloride Chemical compound [Cl-].[Cl-].[Pt+2].[O-][N+](=O)C1=NC=CN1.[O-][N+](=O)C1=NC=CN1 IOSROLCFSUFOFE-UHFFFAOYSA-L 0.000 description 7
- 239000004743 Polypropylene Substances 0.000 description 7
- 102000018120 Recombinases Human genes 0.000 description 7
- 108010091086 Recombinases Proteins 0.000 description 7
- 230000002759 chromosomal effect Effects 0.000 description 7
- 239000013068 control sample Substances 0.000 description 7
- 239000000975 dye Substances 0.000 description 7
- 238000009396 hybridization Methods 0.000 description 7
- 229920001155 polypropylene Polymers 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 238000010561 standard procedure Methods 0.000 description 7
- 230000005945 translocation Effects 0.000 description 7
- 230000032258 transport Effects 0.000 description 7
- 102000003960 Ligases Human genes 0.000 description 6
- 108090000364 Ligases Proteins 0.000 description 6
- 102000006382 Ribonucleases Human genes 0.000 description 6
- 108010083644 Ribonucleases Proteins 0.000 description 6
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 6
- 239000011521 glass Substances 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- 238000011901 isothermal amplification Methods 0.000 description 6
- 230000003278 mimic effect Effects 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- 238000007481 next generation sequencing Methods 0.000 description 6
- 230000008439 repair process Effects 0.000 description 6
- 125000006850 spacer group Chemical group 0.000 description 6
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 5
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 5
- 102000012410 DNA Ligases Human genes 0.000 description 5
- 108010061982 DNA Ligases Proteins 0.000 description 5
- 108060004795 Methyltransferase Proteins 0.000 description 5
- 108700020796 Oncogene Proteins 0.000 description 5
- 238000012408 PCR amplification Methods 0.000 description 5
- 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 5
- 238000013459 approach Methods 0.000 description 5
- 238000003491 array Methods 0.000 description 5
- 238000011109 contamination Methods 0.000 description 5
- 230000007547 defect Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 238000006073 displacement reaction Methods 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 239000000017 hydrogel Substances 0.000 description 5
- 239000011159 matrix material Substances 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 4
- 108700028369 Alleles Proteins 0.000 description 4
- 201000009030 Carcinoma Diseases 0.000 description 4
- 102000010719 DNA-(Apurinic or Apyrimidinic Site) Lyase Human genes 0.000 description 4
- 108010063362 DNA-(Apurinic or Apyrimidinic Site) Lyase Proteins 0.000 description 4
- 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 4
- 201000007224 Myeloproliferative neoplasm Diseases 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 4
- 206010041067 Small cell lung cancer Diseases 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000001574 biopsy Methods 0.000 description 4
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 4
- 230000036425 denaturation Effects 0.000 description 4
- 150000002009 diols Chemical class 0.000 description 4
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 4
- 235000011180 diphosphates Nutrition 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 239000012149 elution buffer Substances 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 4
- 208000032839 leukemia Diseases 0.000 description 4
- 230000000670 limiting effect Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000010899 nucleation Methods 0.000 description 4
- 210000004940 nucleus Anatomy 0.000 description 4
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 238000012175 pyrosequencing Methods 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 208000000587 small cell lung carcinoma Diseases 0.000 description 4
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 210000004243 sweat Anatomy 0.000 description 4
- 210000001138 tear Anatomy 0.000 description 4
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 3
- 229930024421 Adenine Natural products 0.000 description 3
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 3
- 206010061764 Chromosomal deletion Diseases 0.000 description 3
- 208000032170 Congenital Abnormalities Diseases 0.000 description 3
- 206010010356 Congenital anomaly Diseases 0.000 description 3
- 208000000398 DiGeorge Syndrome Diseases 0.000 description 3
- 102000004533 Endonucleases Human genes 0.000 description 3
- 108060002716 Exonuclease Proteins 0.000 description 3
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 3
- 206010025323 Lymphomas Diseases 0.000 description 3
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 3
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 3
- 201000000582 Retinoblastoma Diseases 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 208000026928 Turner syndrome Diseases 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- 229960000643 adenine Drugs 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 238000004132 cross linking Methods 0.000 description 3
- 230000002559 cytogenic effect Effects 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 102000013165 exonuclease Human genes 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000001815 facial effect Effects 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 102000054767 gene variant Human genes 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 230000000977 initiatory effect Effects 0.000 description 3
- 230000036210 malignancy Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 230000036961 partial effect Effects 0.000 description 3
- 150000004713 phosphodiesters Chemical class 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 102000054765 polymorphisms of proteins Human genes 0.000 description 3
- 238000003793 prenatal diagnosis Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000008707 rearrangement Effects 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 210000000582 semen Anatomy 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229940035893 uracil Drugs 0.000 description 3
- 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
- MXHRCPNRJAMMIM-SHYZEUOFSA-N 2'-deoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 MXHRCPNRJAMMIM-SHYZEUOFSA-N 0.000 description 2
- 208000010543 22q11.2 deletion syndrome Diseases 0.000 description 2
- UBKVUFQGVWHZIR-UHFFFAOYSA-N 8-oxoguanine Chemical compound O=C1NC(N)=NC2=NC(=O)N=C21 UBKVUFQGVWHZIR-UHFFFAOYSA-N 0.000 description 2
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 2
- 208000009575 Angelman syndrome Diseases 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- 208000011359 Chromosome disease Diseases 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 230000004544 DNA amplification Effects 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- 230000006820 DNA synthesis Effects 0.000 description 2
- 101100310856 Drosophila melanogaster spri gene Proteins 0.000 description 2
- 208000031448 Genomic Instability Diseases 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 208000026350 Inborn Genetic disease Diseases 0.000 description 2
- 108090001090 Lectins Proteins 0.000 description 2
- 102000004856 Lectins Human genes 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 208000036626 Mental retardation Diseases 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 208000016679 Monosomy X Diseases 0.000 description 2
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 2
- 208000034176 Neoplasms, Germ Cell and Embryonal Diseases 0.000 description 2
- 206010029260 Neuroblastoma Diseases 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 2
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 2
- 201000010769 Prader-Willi syndrome Diseases 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 101150002130 Rb1 gene Proteins 0.000 description 2
- 108050002653 Retinoblastoma protein Proteins 0.000 description 2
- 206010039491 Sarcoma Diseases 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 108010040002 Tumor Suppressor Proteins Proteins 0.000 description 2
- 102000001742 Tumor Suppressor Proteins Human genes 0.000 description 2
- 102000006943 Uracil-DNA Glycosidase Human genes 0.000 description 2
- 108010072685 Uracil-DNA Glycosidase Proteins 0.000 description 2
- 208000036142 Viral infection Diseases 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
- 230000002411 adverse Effects 0.000 description 2
- 238000002669 amniocentesis Methods 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000007698 birth defect Effects 0.000 description 2
- 201000000053 blastoma Diseases 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000003508 chemical denaturation Methods 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 208000024971 chromosomal disease Diseases 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 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 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000003111 delayed 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
- MXHRCPNRJAMMIM-UHFFFAOYSA-N desoxyuridine Natural products C1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 MXHRCPNRJAMMIM-UHFFFAOYSA-N 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 201000008184 embryoma Diseases 0.000 description 2
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 2
- 238000007672 fourth generation sequencing Methods 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 208000016361 genetic disease Diseases 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000009434 installation Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000002523 lectin Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 238000007403 mPCR Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- KVLNTIPUCYZQHA-UHFFFAOYSA-N n-[5-[(2-bromoacetyl)amino]pentyl]prop-2-enamide Chemical compound BrCC(=O)NCCCCCNC(=O)C=C KVLNTIPUCYZQHA-UHFFFAOYSA-N 0.000 description 2
- 102000044158 nucleic acid binding protein Human genes 0.000 description 2
- 108700020942 nucleic acid binding protein Proteins 0.000 description 2
- 239000012038 nucleophile Substances 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 230000005298 paramagnetic effect Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000011009 performance qualification Methods 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000035935 pregnancy Effects 0.000 description 2
- 238000009598 prenatal testing Methods 0.000 description 2
- 229910052761 rare earth metal Inorganic materials 0.000 description 2
- 238000007894 restriction fragment length polymorphism technique Methods 0.000 description 2
- 238000005096 rolling process Methods 0.000 description 2
- 239000012898 sample dilution Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000000527 sonication Methods 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000000153 supplemental effect Effects 0.000 description 2
- 238000001847 surface plasmon resonance imaging Methods 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 235000011178 triphosphate Nutrition 0.000 description 2
- 239000001226 triphosphate Substances 0.000 description 2
- 210000003932 urinary bladder Anatomy 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- JUIKUQOUMZUFQT-UHFFFAOYSA-N 2-bromoacetamide Chemical group NC(=O)CBr JUIKUQOUMZUFQT-UHFFFAOYSA-N 0.000 description 1
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- 208000026817 47,XYY syndrome Diseases 0.000 description 1
- 206010000234 Abortion spontaneous Diseases 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 208000036762 Acute promyelocytic leukaemia Diseases 0.000 description 1
- 102100027211 Albumin Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000003730 Alpha-catenin Human genes 0.000 description 1
- 108090000020 Alpha-catenin Proteins 0.000 description 1
- 101710092462 Alpha-hemolysin Proteins 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 101150111062 C gene Proteins 0.000 description 1
- 208000014392 Cat-eye syndrome Diseases 0.000 description 1
- 206010008805 Chromosomal abnormalities Diseases 0.000 description 1
- 208000031404 Chromosome Aberrations Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 108091028732 Concatemer Proteins 0.000 description 1
- 208000002330 Congenital Heart Defects Diseases 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 108010043471 Core Binding Factor Alpha 2 Subunit Proteins 0.000 description 1
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 230000007018 DNA scission Effects 0.000 description 1
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 description 1
- 230000010777 Disulfide Reduction Effects 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108010051748 Early Growth Response Protein 2 Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102100039788 GTPase NRas Human genes 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 208000021309 Germ cell tumor Diseases 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 101000744505 Homo sapiens GTPase NRas Proteins 0.000 description 1
- 101000638154 Homo sapiens Transmembrane protease serine 2 Proteins 0.000 description 1
- 201000006347 Intellectual Disability Diseases 0.000 description 1
- 208000030979 Language Development disease Diseases 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 208000037699 Monosomy 18p Diseases 0.000 description 1
- 208000019209 Monosomy 22 Diseases 0.000 description 1
- 208000033180 Monosomy 22q13.3 Diseases 0.000 description 1
- 208000010610 Mosaic trisomy 8 Diseases 0.000 description 1
- 101100172630 Mus musculus Eri1 gene Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 241000872931 Myoporum sandwicense Species 0.000 description 1
- 108700026495 N-Myc Proto-Oncogene Proteins 0.000 description 1
- 102100030124 N-myc proto-oncogene protein Human genes 0.000 description 1
- 206010061309 Neoplasm progression Diseases 0.000 description 1
- 208000003019 Neurofibromatosis 1 Diseases 0.000 description 1
- 208000024834 Neurofibromatosis type 1 Diseases 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 208000030852 Parasitic disease Diseases 0.000 description 1
- 108010089430 Phosphoproteins Proteins 0.000 description 1
- 102000007982 Phosphoproteins Human genes 0.000 description 1
- 108010021757 Polynucleotide 5'-Hydroxyl-Kinase Proteins 0.000 description 1
- 102000008422 Polynucleotide 5'-hydroxyl-kinase Human genes 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- 101710086015 RNA ligase Proteins 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 102100025373 Runt-related transcription factor 1 Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 208000019802 Sexually transmitted disease Diseases 0.000 description 1
- 208000020221 Short stature Diseases 0.000 description 1
- 102000005262 Sulfatase Human genes 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical group OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 208000008963 Transient myeloproliferative syndrome Diseases 0.000 description 1
- 102100031989 Transmembrane protease serine 2 Human genes 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 206010062757 Trisomy 15 Diseases 0.000 description 1
- 208000021843 Trisomy 9p Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 206010049644 Williams syndrome Diseases 0.000 description 1
- 208000006254 Wolf-Hirschhorn Syndrome Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 206010056894 XYY syndrome Diseases 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000009056 active transport Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 125000001369 canonical nucleoside group Chemical group 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000004252 chorionic villi Anatomy 0.000 description 1
- 208000004664 chromosome 18p deletion syndrome Diseases 0.000 description 1
- 108091092240 circulating cell-free DNA Proteins 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 208000028831 congenital heart disease Diseases 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 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 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
- 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 1
- 230000006378 damage Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000000326 densiometry Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
- 239000005546 dideoxynucleotide Substances 0.000 description 1
- 239000013024 dilution buffer Substances 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 238000007306 functionalization reaction Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 244000000058 gram-negative pathogen Species 0.000 description 1
- 244000000059 gram-positive pathogen Species 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 239000008241 heterogeneous mixture Substances 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000006607 hypermethylation Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 201000003723 learning disability Diseases 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 230000008376 long-term health Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000533 low birth weight Toxicity 0.000 description 1
- 208000018773 low birth weight Diseases 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 230000036244 malformation Effects 0.000 description 1
- 230000007257 malfunction Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 208000015994 miscarriage Diseases 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000002232 neuromuscular Effects 0.000 description 1
- 238000003499 nucleic acid array Methods 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 230000005257 nucleotidylation Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 210000003254 palate Anatomy 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000009057 passive transport Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 150000008300 phosphoramidites Chemical class 0.000 description 1
- 238000000206 photolithography Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 238000011176 pooling Methods 0.000 description 1
- 238000009609 prenatal screening Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000012521 purified sample Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 108090000850 ribosomal protein S14 Proteins 0.000 description 1
- 102000004314 ribosomal protein S14 Human genes 0.000 description 1
- 210000004708 ribosome subunit Anatomy 0.000 description 1
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 201000000980 schizophrenia Diseases 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000007841 sequencing by ligation Methods 0.000 description 1
- 210000003765 sex chromosome Anatomy 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 231100001055 skeletal defect Toxicity 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 208000000995 spontaneous abortion Diseases 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 108060007951 sulfatase Proteins 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 1
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 208000015344 trisomy 17p Diseases 0.000 description 1
- 208000026485 trisomy X Diseases 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000005751 tumor progression Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000003260 vortexing Methods 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
- C12Q1/6855—Ligating adaptors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1072—Differential gene expression library synthesis, e.g. subtracted libraries, differential screening
-
- 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
- C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
- C12Q2525/10—Modifications characterised by
- C12Q2525/191—Modifications characterised by incorporating an adaptor
-
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
本出願は、その全体が参照により本明細書に組み込まれる、2018年4月2日に出願された米国仮出願第62/651,453号の利益を主張する。
実施形態1.対象から得られた複数の二本鎖鋳型核酸のサンプルを提供することと;
鋳型核酸の両端に汎用アダプターを連結して、複数のアダプター−鋳型−アダプター分子を形成することであって、
複数のアダプター−鋳型−アダプター分子の各々は、汎用アダプターに隣接する鋳型核酸を含み、
汎用アダプターは、二本鎖核酸の領域を含む、形成することと;
第1の汎用プライマー及び第2の汎用プライマーで複数のアダプター−鋳型−アダプター分子を増幅して、増幅されたアダプター−鋳型−アダプター分子を生じさせることと;
増幅されたアダプター−鋳型−アダプター分子の両端から汎用アダプターを除去して、付着していない汎用アダプター及び複数の再生された鋳型核酸(reDNA)を生じさせることと:
を含む方法。
各reDNAは鋳型である、提供することと;
鋳型核酸の両端に汎用アダプターを連結して、複数のアダプター−鋳型−アダプター分子を形成することであって、
複数のアダプター−鋳型−アダプター分子の各々は、汎用アダプターに隣接する鋳型核酸を含み、
汎用アダプターは、(i)二本鎖核酸の領域、及び(ii)少なくとも1つの汎用プライマー結合部位を含む一本鎖非相補的核酸鎖の領域を含み、
それによって、鋳型の少なくとも一部の配列を決定するための配列決定ライブラリーを生成する、形成することと
を含む方法。
増幅部位は、遊離3’末端を有する付着した一本鎖核酸の少なくとも2つの集団を含む、提供することと、
増幅部位を含む表面を、個々のアダプター−鋳型−アダプター分子からのアンプリコンのクローン集団を各々含む複数の増幅部位を産生するのに適した条件下で、複数のアダプター−鋳型−アダプター分子と接触させることと:
をさらに含む、実施形態33〜40のいずれか1つの方法。
a.実施形態1の方法を用いて、複数の再生された鋳型核酸(reDNA)を提供することと;
b.試験サンプル及び工程(a)で得られたreDNAに対して核酸検出試験を行うことと;
c.対照としてreDNAを用いた核酸検出試験の結果を分析することと
を含む、方法。
目的
この実施例は、ヒト血漿を使用する代わりに実験で使用することができる血漿対照材料の作製を記載する。本明細書に記載されるように、このプロセスの間に単離及び精製されるcfDNA入力(reDNA)は、数十のプレートに相当する材料を送達するのに十分に濃縮され、長期間にわたって使用され得る遺伝的に類似したcfDNAの大きなストックを提供する。さらに、使用者は、この調製されたDNAサンプルを血漿希釈物中にスパイクし、ヒト血漿に定量的に類似した代替物を生成し得る。この代替の血漿希釈物は、cfDNAが必要とされる実験においてヒト血漿の品質を模倣するために使用され得、必要とされるより重要な実験のためにヒト血漿を保存する。
このワークフローは、ヒト血漿から抽出されたcfDNA(無細胞DNA)と共に使用するために設計された。cfDNAは、160〜170bpの中央値サイズを有すると予想された。
試薬の調製と取り扱い
ライブラリーは、記載されるように(米国特許第9,323,888号明細書)調製したが、VeriSeq NIPTアダプタープレートをカスタムYアダプターで置き換え、プレートをPCR混合物で溶出した。
Hamilton Star Run Controlを開き、Veriseq NIPT Method(バージョン1.4以降)を、製造者の説明書を使用して実行した。液体検出チェックの後、トラック47上の担体の位置5に溶出緩衝液又はHT1を含む20mLリザーバを除去した。
サンプルプレートをサーモサイクラー上に置き、EPM14プログラムを選択した。EPM 14プログラムの概要を表7に示す。EPM14の完了後、プレートをサーモサイクラーから取り出した。PCRのさらなるサイクルがない場合、次のセクション:「ストレプトアビジン結合、消化、及び精製」に進む。
PCR産物(50μL)を、1.5mL又は2mLチューブ中で950μLの再懸濁緩衝液(RB)と1:20希釈で合わせ、合計1mlとした。ボルテックスミキサーを使用して溶液を混合し、スピンダウンした。
EPM 14生成物(60pg/μLの25uL)を新規96ウェルプレートに添加した。PCRミックス(25uL)をこの溶液に総体積50uLで添加し、混合した。プレートをサーモサイクラー上に置き、表8に概説したEPM 13プログラムを選択した。
このプレートは、後日の使用のために、密封され、2℃〜8℃で保存され得るか、又は密封され、−20℃〜−15℃で保存され得るか、又は一晩以上保存され得る。
試薬の準備と取り扱い
Dynabeads MyOne Streptavidin T1を、室温で最大30分間、ロチッセリー上に置いた。反応は、ディープウェルプレート又は容量の適切なポリプロピレンチューブ中で起こり得る。各消化反応について、十分な消化ミックスを作製するために表9に従う。十分なミックスを作製するのに必要な全反応を体積に乗じる。必要になるまで、SapI消化ミックスを氷上又は4℃に保つ。DNA 1000バイオアナライザー試薬を、暗所内で室温で少なくとも30分間インキュベートする。
消化されるべき各サンプルについて、表10のストレプトアビジンビーズの体積を反応容器に分配した。
洗浄したビーズを有する反応容器を、溶液が透明になるまで(最低1分間)磁石上に置いた。ビーズのみが容器中に残るように、全ての上清を除去した。表10に示すPCR産物の体積を乾燥ビーズに添加し、混合してビーズを再懸濁した。結合ミックスをEppendorf ThermoMixer上で1500 RPM及び20℃で20分間混合した。容器を短時間回転させた後、溶液が透明になるまで(最低1分間)、容器を磁石上に置いた。チューブが磁石上にある間に、全ての上清を除去した。表10に示す消化ミックスの体積を容器に添加し、容器をEppendorf ThermoMixer上に1500 RPM及び37℃で1時間置いた。インキュベーション後、表10の適切な体積のEDTAを反応容器に添加し、混合し、スピンダウンし、溶液が透明になるまで(最低2分間)容器を磁石上に置いた。
試薬の準備と取り扱い
解凍後、10μLのreDNAを10μLの再懸濁緩衝液と合わせることによって、1:2のサンプル希釈のreDNAサンプルのアリコートを作製した。Qubit試薬及びQubit緩衝液を使用して、1:200希釈の色素を作製した。
脱活性化DNA(deDNA)は、任意の酵素活性(連結を含む)を妨げる5’逆方向ddt修飾を有する非ヒト170bp配列由来の増幅DNAである。
1:2サンプル希釈で、reDNA及びdeDNAサンプル当たり2つの別々のチューブ、10μL(高)サンプル入力を有する1つのチューブ、及び5μL(低)入力を有する1つのチューブを使用して、Qubit読み取りを行った。
生成された各900μLの人工血漿は、50%血漿希釈剤(ImmunoChemistry Technologies、Bloomington、MNac)、2mM EDTA、3.5ng reDNA、3.5ng deDNA、及び充填されるdPBSを含んでいた。
reDNAのキャラクタリゼーション
この試験の目的は、reDNA又はreDNA断片が由来する対応するcfDNAのいずれかに対して行われた非侵襲性出生前試験のメトリックを比較することであった。reDNAは、健康な妊娠から得られた全血に由来する血漿から抽出されたcfDNAから、実施例1に記載されるように生成した。血液は、健康な男女の胎児の混合物を含む48の妊娠から得た。適切な同意を得て患者から血液を採取した。
Claims (65)
- 対象から得られた複数の二本鎖鋳型核酸を提供することと;
前記鋳型核酸の両端に汎用アダプターを連結して、前記汎用アダプターに隣接する鋳型核酸を含む複数のアダプター−鋳型−アダプター分子を形成することであって、
前記汎用アダプターは、二本鎖核酸の領域を含む、形成することと;
第1の汎用プライマー及び第2の汎用プライマーで前記複数のアダプター−鋳型−アダプター分子を増幅して、増幅されたアダプター−鋳型−アダプター分子を生じさせることと;
前記増幅されたアダプター−鋳型−アダプター分子の両端から前記汎用アダプターの少なくとも一部を除去して、除去された汎用アダプター及び複数の再生された鋳型核酸(reDNA)を生じさせることと:
を含む方法。 - 前記二本鎖鋳型核酸がDNAである、請求項1に記載の方法。
- 前記二本鎖鋳型核酸が無細胞DNA(cfDNA)を含む、請求項1に記載の方法。
- 前記対象が妊娠しているヒトであり、前記二本鎖鋳型核酸が胎児及び母体核酸の混合物を含む、請求項1に記載の方法。
- 前記複数の二本鎖鋳型核酸がcfDNAを含む、請求項4に記載の方法。
- 前記胎児が遺伝的状態を含まない、請求項4に記載の方法。
- 前記胎児が遺伝的状態を含む、請求項4に記載の方法。
- 前記遺伝的状態が異数性である、請求項6又は7に記載の方法。
- 前記異数性がトリソミーである、請求項8に記載の方法。
- 前記トリソミーが、21トリソミー、18トリソミー、13トリソミー、9トリソミー、8トリソミー、22トリソミー、XXX、XXY、又はXYYである、請求項9に記載の方法。
- 前記遺伝的状態がヌクレオチド配列の過小発現を含む、請求項9又は10に記載の方法。
- 前記対象が新生物を有することが疑われる、請求項1に記載の方法。
- 前記複数の二本鎖鋳型核酸が、循環腫瘍DNA及び無細胞正常DNAを含む、請求項12に記載の方法。
- 前記複数の二本鎖鋳型核酸が、血液、尿、痰、又は便を含む、請求項1に記載の方法。
- 前記連結の前に、前記複数の二本鎖鋳型核酸を処理して、平滑末端になるように末端を修飾することをさらに含む、請求項1に記載の方法。
- 前記連結の前に、前記複数の二本鎖鋳型核酸を処理して、3’末端を修飾して3’突出構造として終結させることをさらに含む、請求項1に記載の方法。
- 前記汎用アダプターが少なくとも1つの汎用プライマー結合部位を含む一本鎖非相補的核酸鎖の領域をさらに含み、前記連結が、前記汎用アダプターの二本鎖核酸の前記領域を前記鋳型核酸の各末端に共有結合的に付着させる、請求項1に記載の方法。
- 前記増幅が指数関数的増幅反応を含む、請求項1に記載の方法。
- 前記指数関数的増幅反応がポリメラーゼ連鎖反応(PCR)を含む、請求項18に記載の方法。
- 前記増幅が、低いエラー率を有するDNAポリメラーゼを含む、請求項1に記載の方法。
- 前記汎用アダプターが制限エンドヌクレアーゼ認識部位を含み、前記除去することが、増幅されたアダプター−鋳型−アダプター分子を制限エンドヌクレアーゼに曝露すること、及び前記アダプター−鋳型−アダプター分子を切断して、除去された汎用アダプター及び複数のreDNAを生じさせることを含む、請求項1に記載の方法。
- 前記制限エンドヌクレアーゼの前記切断部位及び前記認識部位が別々である、請求項21に記載の方法。
- 前記制限エンドヌクレアーゼがSapI、MlyI、又はBpuEIである、請求項22に記載の方法。
- 前記reDNAが、前記鋳型の各末端に前記汎用アダプターの一部を保持する、請求項1に記載の方法。
- 前記reDNAが、前記鋳型の各末端に前記汎用アダプターの一部を保持しない、請求項1に記載の方法。
- 前記第1の汎用プライマー及び前記第2の汎用プライマーが捕捉剤を含む、請求項1に記載の方法。
- 前記捕捉剤がビオチンを含む、請求項26に記載の方法。
- 前記汎用アダプターが制限エンドヌクレアーゼ認識部位を含み、前記除去することが、前記アダプター−鋳型−アダプター分子を、前記リガンドに結合して、結合された増幅されたアダプター−鋳型−アダプター分子を生じさせる化合物を含む表面と接触させることと、前記結合された増幅されたアダプター−鋳型−アダプター分子を、前記結合された増幅されたアダプター−鋳型−アダプター分子を切断して除去された汎用アダプター及び複数のreDNAを生じさせる制限エンドヌクレアーゼに曝露することとを含み、前記除去された汎用アダプターが前記表面に結合される、請求項26に記載の方法。
- 前記除去することが、前記除去された汎用アダプターを前記reDNAから分離することをさらに含む、請求項26に記載の方法。
- 前記分離することが、前記除去された汎用アダプター及び前記reDNAを含む混合物を、前記リガンドに結合する化合物と接触させることを含み、前記化合物が表面に付着され、前記除去された汎用アダプターが前記表面に結合される、請求項29に記載の方法。
- 前記表面がビーズを含む、請求項28又は30に記載の方法。
- 前記結合された除去された汎用アダプターを含む前記表面を除去して、前記除去された汎用アダプターを前記reDNAから分離することをさらに含む、請求項28又は30に記載の方法。
- 前記除去することが、所定のサイズ範囲内に入るreDNAの選択を含む、請求項1に記載の方法。
- 対象から得られたサンプルに由来する複数の再生鋳型核酸(reDNA)を提供することであって、
各reDNAは鋳型である、提供することと;
前記鋳型核酸の両端に汎用アダプターを連結して、前記汎用アダプターに隣接する鋳型核酸を含む複数のアダプター−鋳型−アダプター分子を形成することであって、
前記汎用アダプターは、(i)二本鎖核酸の領域、及び(ii)少なくとも1つの汎用プライマー結合部位を含む一本鎖非相補的核酸鎖の領域を含み、
それによって、鋳型の少なくとも一部の配列を決定するための配列決定ライブラリーを生成する、形成することと
を含む方法。 - 一本鎖非相補的核酸鎖の前記領域が、少なくとも1つの汎用伸長プライマー結合部位をさらに含む、請求項34に記載の方法。
- 一本鎖非相補的核酸鎖の前記領域に対して遠位の二本鎖核酸の前記領域が平滑末端構造として終結する、請求項34に記載の方法。
- 前記複数の鋳型が平滑末端構造を含む、請求項36に記載の方法。
- 一本鎖非相補的核酸鎖の前記領域に対して遠位の二本鎖核酸の前記領域が、3’突出構造として終結する、請求項34に記載の方法。
- 前記3’突出構造が、1〜4ヌクレオチドの突出構造を含む、請求項38に記載の方法。
- 前記3’突出構造がTヌクレオチドの突出を含む、請求項38に記載の方法。
- 前記鋳型が、二本鎖核酸の前記領域の前記3’突出構造に相補的な3’突出構造を含む、請求項38に記載の方法。
- 複数の増幅部位を含む表面を提供することであって、
前記増幅部位は、遊離3’末端を有する付着した一本鎖核酸の少なくとも2つの集団を含む、提供することと、
増幅部位を含む前記表面を、個々のアダプター−鋳型−アダプター分子からのアンプリコンのクローン集団を各々含む複数の増幅部位を産生するのに適した条件下で、前記複数のアダプター−鋳型−アダプター分子と接触させることと、
をさらに含む、請求項34に記載の方法。 - 前記複数のアダプター−鋳型−アダプター分子の数が増幅部位の数を超え、前記アダプター−鋳型−アダプター分子が前記増幅部位への流体アクセスを有し、前記増幅部位の各々がいくつかのアダプター−鋳型−アダプター分子のための容量を含む、請求項42に記載の方法。
- 前記接触させることが、(i)前記アダプター−鋳型−アダプター分子を平均輸送速度で前記増幅部位に輸送すること、及び(ii)前記増幅部位にある前記アダプター−鋳型−アダプター分子を平均増幅速度で増幅することを同時に含み、前記平均増幅速度が前記平均輸送速度を超える、請求項42に記載の方法。
- 前記汎用アダプターが制限エンドヌクレアーゼ認識部位を含む、請求項1に記載のアダプター−鋳型−アダプター分子を含む組成物。
- 前記制限エンドヌクレアーゼがSapI、MlyI、又はBpuEIである、請求項45に記載の組成物。
- 人工生物学的流体をさらに含む、請求項1に記載の方法によって生成された前記複数のreDNAを含む組成物。
- 前記人工生物学的流体が人工血漿を含む、請求項47に記載の組成物。
- 前記対象が病原体感染を有することが疑われる、請求項1に記載の方法。
- 前記病原体が細菌病原体又はウイルス病原体である、請求項49に記載の方法。
- 前記サンプルがウイルスDNA又は細菌DNAを含む、請求項50に記載の方法。
- 前記サンプルが、血液、尿、痰、又は便を含む、請求項50に記載の方法。
- 核酸検出試験において対照を使用する方法であって:
a.請求項1に記載の方法を用いて、複数の再生された鋳型核酸(reDNA)を提供することと;
b.試験サンプル及び工程(a)で得られた前記reDNAに対して核酸検出試験を行うことと;
c.対照として前記reDNAを用いた前記核酸検出試験の結果を分析することと
を含む、方法。 - 前記核酸検出試験が配列決定を含む、請求項53に記載の方法。
- 前記核酸検出試験がマイクロアレイ分析を含む、請求項53に記載の方法。
- 前記核酸検出試験が酵素プロセスを含む、請求項53に記載の方法。
- 請求項1に記載の方法を用いて得られた複数の再生鋳型核酸(reDNA)に対して核酸検出試験を実施することを含む方法。
- 前記reDNAが、ライブラリー調製方法の品質のための対照である、請求項57に記載の方法。
- 前記reDNAが、配列決定器具のための較正対照である、請求項57に記載の方法。
- 前記reDNAが、アレイ機器のための較正対照である、請求項57に記載の方法。
- 前記reDNAが、核酸配列決定試験のための検証対照である、請求項57に記載の方法。
- 前記reDNAが、配列決定に基づく非侵襲性出生前試験のための検証対照である、請求項61に記載の方法。
- 前記reDNAが、配列決定に基づく腫瘍検出試験のための検証対照である、請求項61に記載の方法。
- 前記reDNAが、配列決定に基づくコンパニオン診断試験のための検証対照である、請求項61に記載の方法。
- 前記コンパニオン診断試験が、処置レジメンにおいて治療処置が適切であるかどうかを決定するために、サンプル中の遺伝子変異体の存在又は同一性を決定する、請求項64に記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862651453P | 2018-04-02 | 2018-04-02 | |
US62/651,453 | 2018-04-02 | ||
PCT/US2019/025304 WO2019195225A1 (en) | 2018-04-02 | 2019-04-02 | Compositions and methods for making controls for sequence-based genetic testing |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2020524488A true JP2020524488A (ja) | 2020-08-20 |
JP6974504B2 JP6974504B2 (ja) | 2021-12-01 |
Family
ID=66223832
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2019566955A Active JP6974504B2 (ja) | 2018-04-02 | 2019-04-02 | 配列に基づく遺伝子試験のための対照を作製するための組成物及び方法 |
Country Status (8)
Country | Link |
---|---|
US (1) | US20190367909A1 (ja) |
EP (1) | EP3775278A1 (ja) |
JP (1) | JP6974504B2 (ja) |
KR (1) | KR102383799B1 (ja) |
CN (1) | CN110832086A (ja) |
AU (1) | AU2019248635B2 (ja) |
CA (1) | CA3064622A1 (ja) |
WO (1) | WO2019195225A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111500696B (zh) * | 2020-03-18 | 2023-03-14 | 中国科学院苏州生物医学工程技术研究所 | 基于飞行时间质谱检测孕妇外周血游离rna筛查21-三体综合征的试剂盒 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050202490A1 (en) * | 2004-03-08 | 2005-09-15 | Makarov Vladimir L. | Methods and compositions for generating and amplifying DNA libraries for sensitive detection and analysis of DNA methylation |
US20130029852A1 (en) * | 2010-01-19 | 2013-01-31 | Verinata Health, Inc. | Detecting and classifying copy number variation |
JP2014223089A (ja) * | 2008-04-25 | 2014-12-04 | 地方独立行政法人東京都健康長寿医療センター | Dnaメチル化分析方法 |
Family Cites Families (78)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4683195A (en) | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
CA1323293C (en) | 1987-12-11 | 1993-10-19 | Keith C. Backman | Assay using template-dependent nucleic acid probe reorganization |
CA1341584C (en) | 1988-04-06 | 2008-11-18 | Bruce Wallace | Method of amplifying and detecting nucleic acid sequences |
WO1989009835A1 (en) | 1988-04-08 | 1989-10-19 | The Salk Institute For Biological Studies | Ligase-based amplification method |
US5130238A (en) | 1988-06-24 | 1992-07-14 | Cangene Corporation | Enhanced nucleic acid amplification process |
JP2801051B2 (ja) | 1988-06-24 | 1998-09-21 | アムジエン・インコーポレーテツド | 核酸塩基配列を検出するための方法及び試薬 |
WO1990001069A1 (en) | 1988-07-20 | 1990-02-08 | Segev Diagnostics, Inc. | Process for amplifying and detecting nucleic acid sequences |
US5185243A (en) | 1988-08-25 | 1993-02-09 | Syntex (U.S.A.) Inc. | Method for detection of specific nucleic acid sequences |
CA2044616A1 (en) | 1989-10-26 | 1991-04-27 | Roger Y. Tsien | Dna sequencing |
US5573907A (en) | 1990-01-26 | 1996-11-12 | Abbott Laboratories | Detecting and amplifying target nucleic acids using exonucleolytic activity |
AU635105B2 (en) | 1990-01-26 | 1993-03-11 | Abbott Laboratories | Improved method of amplifying target nucleic acids applicable to both polymerase and ligase chain reactions |
US5223414A (en) | 1990-05-07 | 1993-06-29 | Sri International | Process for nucleic acid hybridization and amplification |
US5455166A (en) | 1991-01-31 | 1995-10-03 | Becton, Dickinson And Company | Strand displacement amplification |
JP3175110B2 (ja) | 1994-02-07 | 2001-06-11 | オーキッド・バイオサイエンシーズ・インコーポレイテッド | リガーゼ/ポリメラーゼ媒体された単一ヌクレオチド多型のジェネティックビットアナリシスおよび遺伝子解析におけるその使用 |
AU687535B2 (en) | 1994-03-16 | 1998-02-26 | Gen-Probe Incorporated | Isothermal strand displacement nucleic acid amplification |
US5846719A (en) | 1994-10-13 | 1998-12-08 | Lynx Therapeutics, Inc. | Oligonucleotide tags for sorting and identification |
US5750341A (en) | 1995-04-17 | 1998-05-12 | Lynx Therapeutics, Inc. | DNA sequencing by parallel oligonucleotide extensions |
GB9620209D0 (en) | 1996-09-27 | 1996-11-13 | Cemu Bioteknik Ab | Method of sequencing DNA |
GB9626815D0 (en) | 1996-12-23 | 1997-02-12 | Cemu Bioteknik Ab | Method of sequencing DNA |
ES2563643T3 (es) | 1997-04-01 | 2016-03-15 | Illumina Cambridge Limited | Método de secuenciación de ácido nucleico |
US6969488B2 (en) | 1998-05-22 | 2005-11-29 | Solexa, Inc. | System and apparatus for sequential processing of analytes |
AR021833A1 (es) | 1998-09-30 | 2002-08-07 | Applied Research Systems | Metodos de amplificacion y secuenciacion de acido nucleico |
US6274320B1 (en) | 1999-09-16 | 2001-08-14 | Curagen Corporation | Method of sequencing a nucleic acid |
US7955794B2 (en) | 2000-09-21 | 2011-06-07 | Illumina, Inc. | Multiplex nucleic acid reactions |
US7582420B2 (en) | 2001-07-12 | 2009-09-01 | Illumina, Inc. | Multiplex nucleic acid reactions |
US7611869B2 (en) | 2000-02-07 | 2009-11-03 | Illumina, Inc. | Multiplexed methylation detection methods |
US7001792B2 (en) | 2000-04-24 | 2006-02-21 | Eagle Research & Development, Llc | Ultra-fast nucleic acid sequencing device and a method for making and using the same |
CA2415897A1 (en) | 2000-07-07 | 2002-01-17 | Susan H. Hardin | Real-time sequence determination |
EP1354064A2 (en) | 2000-12-01 | 2003-10-22 | Visigen Biotechnologies, Inc. | Enzymatic nucleic acid synthesis: compositions and methods for altering monomer incorporation fidelity |
AR031640A1 (es) | 2000-12-08 | 2003-09-24 | Applied Research Systems | Amplificacion isotermica de acidos nucleicos en un soporte solido |
US7057026B2 (en) | 2001-12-04 | 2006-06-06 | Solexa Limited | Labelled nucleotides |
US7399590B2 (en) | 2002-02-21 | 2008-07-15 | Asm Scientific, Inc. | Recombinase polymerase amplification |
US8030000B2 (en) | 2002-02-21 | 2011-10-04 | Alere San Diego, Inc. | Recombinase polymerase amplification |
DK3363809T3 (da) | 2002-08-23 | 2020-05-04 | Illumina Cambridge Ltd | Modificerede nukleotider til polynukleotidsekvensering |
CA2498764C (en) | 2002-09-20 | 2015-11-10 | New England Biolabs, Inc. | Helicase dependent amplification of nucleic acids |
WO2005003304A2 (en) | 2003-06-20 | 2005-01-13 | Illumina, Inc. | Methods and compositions for whole genome amplification and genotyping |
GB0321306D0 (en) | 2003-09-11 | 2003-10-15 | Solexa Ltd | Modified polymerases for improved incorporation of nucleotide analogues |
EP1701785A1 (en) | 2004-01-07 | 2006-09-20 | Solexa Ltd. | Modified molecular arrays |
CN101914620B (zh) | 2004-09-17 | 2014-02-12 | 加利福尼亚太平洋生命科学公司 | 核酸测序的方法 |
EP1828412B2 (en) | 2004-12-13 | 2019-01-09 | Illumina Cambridge Limited | Improved method of nucleotide detection |
JP4990886B2 (ja) | 2005-05-10 | 2012-08-01 | ソレックサ リミテッド | 改良ポリメラーゼ |
GB0514936D0 (en) | 2005-07-20 | 2005-08-24 | Solexa Ltd | Preparation of templates for nucleic acid sequencing |
US7405281B2 (en) | 2005-09-29 | 2008-07-29 | Pacific Biosciences Of California, Inc. | Fluorescent nucleotide analogs and uses therefor |
GB0522310D0 (en) | 2005-11-01 | 2005-12-07 | Solexa Ltd | Methods of preparing libraries of template polynucleotides |
US20070172839A1 (en) * | 2006-01-24 | 2007-07-26 | Smith Douglas R | Asymmetrical adapters and methods of use thereof |
EP2423334A3 (en) | 2006-02-02 | 2012-04-18 | The Board of Trustees of The Leland Stanford Junior University | Non-invasive fetal genetic screening by digital analysis |
EP2021503A1 (en) | 2006-03-17 | 2009-02-11 | Solexa Ltd. | Isothermal methods for creating clonal single molecule arrays |
US8241573B2 (en) | 2006-03-31 | 2012-08-14 | Illumina, Inc. | Systems and devices for sequence by synthesis analysis |
WO2008051530A2 (en) | 2006-10-23 | 2008-05-02 | Pacific Biosciences Of California, Inc. | Polymerase enzymes and reagents for enhanced nucleic acid sequencing |
US8349167B2 (en) | 2006-12-14 | 2013-01-08 | Life Technologies Corporation | Methods and apparatus for detecting molecular interactions using FET arrays |
ES2923759T3 (es) | 2006-12-14 | 2022-09-30 | Life Technologies Corp | Aparato para medir analitos utilizando matrices de FET |
US8262900B2 (en) | 2006-12-14 | 2012-09-11 | Life Technologies Corporation | Methods and apparatus for measuring analytes using large scale FET arrays |
EP2121983A2 (en) | 2007-02-02 | 2009-11-25 | Illumina Cambridge Limited | Methods for indexing samples and sequencing multiple nucleotide templates |
WO2008096146A1 (en) * | 2007-02-07 | 2008-08-14 | Solexa Limited | Preparation of templates for methylation analysis |
EA028642B1 (ru) | 2007-07-23 | 2017-12-29 | Те Чайниз Юниверсити Ов Гонгконг | Способ пренатальной диагностики фетальной хромосомной анэуплоидии |
WO2010003132A1 (en) | 2008-07-02 | 2010-01-07 | Illumina Cambridge Ltd. | Using populations of beads for the fabrication of arrays on surfaces |
US20100137143A1 (en) | 2008-10-22 | 2010-06-03 | Ion Torrent Systems Incorporated | Methods and apparatus for measuring analytes |
US9323888B2 (en) | 2010-01-19 | 2016-04-26 | Verinata Health, Inc. | Detecting and classifying copy number variation |
US20120270739A1 (en) | 2010-01-19 | 2012-10-25 | Verinata Health, Inc. | Method for sample analysis of aneuploidies in maternal samples |
US8951781B2 (en) | 2011-01-10 | 2015-02-10 | Illumina, Inc. | Systems, methods, and apparatuses to image a sample for biological or chemical analysis |
WO2012170936A2 (en) | 2011-06-09 | 2012-12-13 | Illumina, Inc. | Patterned flow-cells useful for nucleic acid analysis |
WO2013044018A1 (en) | 2011-09-23 | 2013-03-28 | Illumina, Inc. | Methods and compositions for nucleic acid sequencing |
CA3003082C (en) | 2011-10-28 | 2020-12-15 | Illumina, Inc. | Microarray fabrication system and method |
US8938309B2 (en) | 2012-01-16 | 2015-01-20 | Greatbatch Ltd. | Elevated hermetic feedthrough insulator adapted for side attachment of electrical conductors on the body fluid side of an active implantable medical device |
EP4219012A1 (en) | 2012-04-03 | 2023-08-02 | Illumina, Inc. | Method of imaging a substrate comprising fluorescent features and use of the method in nucleic acid sequencing |
US8895249B2 (en) | 2012-06-15 | 2014-11-25 | Illumina, Inc. | Kinetic exclusion amplification of nucleic acid libraries |
WO2014014497A1 (en) * | 2012-07-20 | 2014-01-23 | Verinata Health, Inc. | Detecting and classifying copy number variation in a cancer genome |
WO2014028778A1 (en) * | 2012-08-15 | 2014-02-20 | Natera, Inc. | Methods and compositions for reducing genetic library contamination |
US9512422B2 (en) | 2013-02-26 | 2016-12-06 | Illumina, Inc. | Gel patterned surfaces |
AU2014233373B2 (en) * | 2013-03-15 | 2019-10-24 | Verinata Health, Inc. | Generating cell-free DNA libraries directly from blood |
DK3017065T3 (en) | 2013-07-01 | 2018-11-26 | Illumina Inc | Catalyst-free Surface functionalization and polymer grafting |
US9677132B2 (en) | 2014-01-16 | 2017-06-13 | Illumina, Inc. | Polynucleotide modification on solid support |
PL3212684T3 (pl) | 2014-10-31 | 2020-10-19 | Illumina Cambridge Limited | Polimery i powłoki z kopolimeru DNA |
EP3725893A1 (en) | 2015-02-10 | 2020-10-21 | Illumina, Inc. | Compositions for analyzing cellular components |
US10095831B2 (en) | 2016-02-03 | 2018-10-09 | Verinata Health, Inc. | Using cell-free DNA fragment size to determine copy number variations |
CN109844132B (zh) * | 2016-08-10 | 2023-11-03 | 格瑞尔有限责任公司 | 分析核酸片段的方法 |
CA3207879A1 (en) * | 2017-01-24 | 2018-08-02 | Sequenom, Inc. | Methods and processes for assessment of genetic variations |
-
2019
- 2019-04-02 CA CA3064622A patent/CA3064622A1/en active Pending
- 2019-04-02 EP EP19718517.6A patent/EP3775278A1/en active Pending
- 2019-04-02 AU AU2019248635A patent/AU2019248635B2/en active Active
- 2019-04-02 CN CN201980002978.2A patent/CN110832086A/zh active Pending
- 2019-04-02 JP JP2019566955A patent/JP6974504B2/ja active Active
- 2019-04-02 KR KR1020197037047A patent/KR102383799B1/ko active IP Right Grant
- 2019-04-02 WO PCT/US2019/025304 patent/WO2019195225A1/en unknown
- 2019-04-02 US US16/372,622 patent/US20190367909A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050202490A1 (en) * | 2004-03-08 | 2005-09-15 | Makarov Vladimir L. | Methods and compositions for generating and amplifying DNA libraries for sensitive detection and analysis of DNA methylation |
JP2014223089A (ja) * | 2008-04-25 | 2014-12-04 | 地方独立行政法人東京都健康長寿医療センター | Dnaメチル化分析方法 |
US20130029852A1 (en) * | 2010-01-19 | 2013-01-31 | Verinata Health, Inc. | Detecting and classifying copy number variation |
Non-Patent Citations (1)
Title |
---|
SIDDHARTH S DEY: "Integrated genome and transcriptome sequencing of the same cell", NATURE BIOTECHNOLOGY, vol. 33, JPN6020046625, 2015, pages 285 - 289, XP055588393, ISSN: 0004518785, DOI: 10.1038/nbt.3129 * |
Also Published As
Publication number | Publication date |
---|---|
AU2019248635B2 (en) | 2022-01-27 |
JP6974504B2 (ja) | 2021-12-01 |
US20190367909A1 (en) | 2019-12-05 |
WO2019195225A1 (en) | 2019-10-10 |
CN110832086A (zh) | 2020-02-21 |
KR102383799B1 (ko) | 2022-04-05 |
EP3775278A1 (en) | 2021-02-17 |
AU2019248635A1 (en) | 2019-12-12 |
KR20200005658A (ko) | 2020-01-15 |
CA3064622A1 (en) | 2019-10-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
RU2770879C2 (ru) | Полногеномные библиотеки отдельных клеток для бисульфитного секвенирования | |
KR102475710B1 (ko) | 단일 세포 전체 게놈 라이브러리 및 이의 제조를 위한 조합 인덱싱 방법 | |
US20190024141A1 (en) | Direct Capture, Amplification and Sequencing of Target DNA Using Immobilized Primers | |
JP2020533272A (ja) | 低減した増幅バイアスによるハイスループット単一細胞シークエンシング | |
JP2022036975A (ja) | ナノポア技術を用いた短いdna断片の迅速な配列決定 | |
EP3612641A1 (en) | Compositions and methods for library construction and sequence analysis | |
CN110914449A (zh) | 构建测序文库 | |
JP6974504B2 (ja) | 配列に基づく遺伝子試験のための対照を作製するための組成物及び方法 | |
US20230112730A1 (en) | Compositions and methods for oncology precision assays |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20191220 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20201207 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210308 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210607 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210907 |
|
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: 20211004 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20211104 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6974504 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |