JP6930910B2 - Rna転写産物バリアントを定量するための方法及び製品 - Google Patents
Rna転写産物バリアントを定量するための方法及び製品 Download PDFInfo
- Publication number
- JP6930910B2 JP6930910B2 JP2017501008A JP2017501008A JP6930910B2 JP 6930910 B2 JP6930910 B2 JP 6930910B2 JP 2017501008 A JP2017501008 A JP 2017501008A JP 2017501008 A JP2017501008 A JP 2017501008A JP 6930910 B2 JP6930910 B2 JP 6930910B2
- Authority
- JP
- Japan
- Prior art keywords
- artificial
- sequence
- family
- transcript
- molecules
- 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 99
- 150000007523 nucleic acids Chemical class 0.000 claims description 197
- 108020004707 nucleic acids Proteins 0.000 claims description 195
- 102000039446 nucleic acids Human genes 0.000 claims description 195
- 108090000623 proteins and genes Proteins 0.000 claims description 78
- 239000002773 nucleotide Substances 0.000 claims description 52
- 125000003729 nucleotide group Chemical group 0.000 claims description 52
- 238000012163 sequencing technique Methods 0.000 claims description 52
- 238000013518 transcription Methods 0.000 claims description 43
- 230000035897 transcription Effects 0.000 claims description 43
- 108020004414 DNA Proteins 0.000 claims description 36
- 238000011002 quantification Methods 0.000 claims description 35
- 108700024394 Exon Proteins 0.000 claims description 29
- 108700005078 Synthetic Genes Proteins 0.000 claims description 24
- 108020004705 Codon Proteins 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 108700009124 Transcription Initiation Site Proteins 0.000 claims description 4
- 230000005030 transcription termination Effects 0.000 claims description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 3
- 210000002784 stomach Anatomy 0.000 claims 1
- 239000000523 sample Substances 0.000 description 79
- 108020004999 messenger RNA Proteins 0.000 description 41
- 239000000203 mixture Substances 0.000 description 38
- 238000005259 measurement Methods 0.000 description 36
- 241000282414 Homo sapiens Species 0.000 description 25
- 238000007481 next generation sequencing Methods 0.000 description 23
- 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 22
- 238000002360 preparation method Methods 0.000 description 22
- 238000002474 experimental method Methods 0.000 description 19
- 238000003753 real-time PCR Methods 0.000 description 19
- 239000003795 chemical substances by application Substances 0.000 description 18
- 230000000692 anti-sense effect Effects 0.000 description 16
- 238000001514 detection method Methods 0.000 description 16
- 238000003559 RNA-seq method Methods 0.000 description 15
- 238000009826 distribution Methods 0.000 description 15
- 239000013612 plasmid Substances 0.000 description 14
- 108091028043 Nucleic acid sequence Proteins 0.000 description 12
- 108010029485 Protein Isoforms Proteins 0.000 description 12
- 102000001708 Protein Isoforms Human genes 0.000 description 12
- 239000013614 RNA sample Substances 0.000 description 12
- 238000013507 mapping Methods 0.000 description 12
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 11
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 11
- 108091036407 Polyadenylation Proteins 0.000 description 11
- 229940029575 guanosine Drugs 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 238000000746 purification Methods 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 10
- 238000004422 calculation algorithm Methods 0.000 description 10
- 238000012937 correction Methods 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 10
- 230000014509 gene expression Effects 0.000 description 9
- 238000012545 processing Methods 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 108020005067 RNA Splice Sites Proteins 0.000 description 8
- 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 8
- 210000004556 brain Anatomy 0.000 description 8
- 230000000977 initiatory effect Effects 0.000 description 8
- 239000002679 microRNA Substances 0.000 description 8
- 241000206602 Eukaryota Species 0.000 description 7
- 108700011259 MicroRNAs Proteins 0.000 description 7
- 108091034057 RNA (poly(A)) Proteins 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000012634 fragment Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 108020005544 Antisense RNA Proteins 0.000 description 6
- 101100184147 Caenorhabditis elegans mix-1 gene Proteins 0.000 description 6
- 108091027974 Mature messenger RNA Proteins 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 230000000295 complement effect Effects 0.000 description 6
- 238000001035 drying Methods 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 238000010208 microarray analysis Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000003381 stabilizer Substances 0.000 description 6
- 230000003612 virological effect Effects 0.000 description 6
- 108091092195 Intron Proteins 0.000 description 5
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 5
- 150000003838 adenosines Chemical class 0.000 description 5
- 238000000546 chi-square test Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 230000009467 reduction Effects 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 238000004088 simulation Methods 0.000 description 5
- 241001515965 unidentified phage Species 0.000 description 5
- 238000010200 validation analysis Methods 0.000 description 5
- 108091026821 Artificial microRNA Proteins 0.000 description 4
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 4
- 230000006820 DNA synthesis Effects 0.000 description 4
- 102000016911 Deoxyribonucleases Human genes 0.000 description 4
- 108010053770 Deoxyribonucleases Proteins 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 101710137500 T7 RNA polymerase Proteins 0.000 description 4
- 229960005305 adenosine Drugs 0.000 description 4
- 239000011324 bead Substances 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 230000007717 exclusion Effects 0.000 description 4
- 238000002493 microarray Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000009827 uniform distribution Methods 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 102000053602 DNA Human genes 0.000 description 3
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 3
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 108700005081 Overlapping Genes Proteins 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 238000004590 computer program Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000009396 hybridization Methods 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000008488 polyadenylation Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000006641 stabilisation Effects 0.000 description 3
- 238000011105 stabilization Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 108020003215 DNA Probes Proteins 0.000 description 2
- 239000003298 DNA probe Substances 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 102100036498 Dual specificity testis-specific protein kinase 2 Human genes 0.000 description 2
- 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 2
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 2
- 102100039335 HAUS augmin-like complex subunit 5 Human genes 0.000 description 2
- 101000714156 Homo sapiens Dual specificity testis-specific protein kinase 2 Proteins 0.000 description 2
- 101001035821 Homo sapiens HAUS augmin-like complex subunit 5 Proteins 0.000 description 2
- 101001091379 Homo sapiens Kallikrein-5 Proteins 0.000 description 2
- 101001130147 Homo sapiens Probable D-lactate dehydrogenase, mitochondrial Proteins 0.000 description 2
- 101000671649 Homo sapiens Upstream stimulatory factor 2 Proteins 0.000 description 2
- 102100034868 Kallikrein-5 Human genes 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 108700001094 Plant Genes Proteins 0.000 description 2
- 102100031708 Probable D-lactate dehydrogenase, mitochondrial Human genes 0.000 description 2
- 102000006382 Ribonucleases Human genes 0.000 description 2
- 108010083644 Ribonucleases Proteins 0.000 description 2
- 102100040103 Upstream stimulatory factor 2 Human genes 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000000205 computational method Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 238000011157 data evaluation Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 108091070501 miRNA Proteins 0.000 description 2
- 238000003203 nucleic acid sequencing method Methods 0.000 description 2
- 230000037452 priming Effects 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 239000003161 ribonuclease inhibitor Substances 0.000 description 2
- 108020004418 ribosomal RNA Proteins 0.000 description 2
- 238000005204 segregation Methods 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- 210000001324 spliceosome Anatomy 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000005026 transcription initiation Effects 0.000 description 2
- 230000002103 transcriptional effect Effects 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
- GNXFOGHNGIVQEH-UHFFFAOYSA-N 2-hydroxy-3-(2-methoxyphenoxy)propyl carbamate Chemical compound COC1=CC=CC=C1OCC(O)COC(N)=O GNXFOGHNGIVQEH-UHFFFAOYSA-N 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 108700020462 BRCA2 Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 101150008921 Brca2 gene Proteins 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 102000001493 Cyclophilins Human genes 0.000 description 1
- 108010068682 Cyclophilins Proteins 0.000 description 1
- 238000000018 DNA microarray Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 101000878605 Homo sapiens Low affinity immunoglobulin epsilon Fc receptor Proteins 0.000 description 1
- 101000655352 Homo sapiens Telomerase reverse transcriptase Proteins 0.000 description 1
- 238000009015 Human TaqMan MicroRNA Assay kit Methods 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 101710203526 Integrase Proteins 0.000 description 1
- 102100038007 Low affinity immunoglobulin epsilon Fc receptor Human genes 0.000 description 1
- 241001599018 Melanogaster Species 0.000 description 1
- 241000203407 Methanocaldococcus jannaschii Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 101100202339 Mus musculus Slc6a13 gene Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000772415 Neovison vison Species 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 241000209094 Oryza Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000271578 Ovophis okinavensis Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 241000015864 Protobothrops flavoviridis Species 0.000 description 1
- 238000013381 RNA quantification Methods 0.000 description 1
- 101100202330 Rattus norvegicus Slc6a11 gene Proteins 0.000 description 1
- 101100273253 Rhizopus niveus RNAP gene Proteins 0.000 description 1
- 102000042773 Small Nucleolar RNA Human genes 0.000 description 1
- 108020003224 Small Nucleolar RNA Proteins 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 108091023045 Untranslated Region Proteins 0.000 description 1
- 206010046865 Vaccinia virus infection Diseases 0.000 description 1
- 241000269457 Xenopus tropicalis Species 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004847 absorption spectroscopy Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N adenyl group Chemical group N1=CN=C2N=CNC2=C1N GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000000712 assembly Effects 0.000 description 1
- 238000000429 assembly Methods 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 101150049515 bla gene Proteins 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000009274 differential gene expression Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 235000003869 genetically modified organism Nutrition 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 238000009499 grossing Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000009830 intercalation Methods 0.000 description 1
- 101150066555 lacZ gene Proteins 0.000 description 1
- 235000019689 luncheon sausage Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000011807 nanoball Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 210000000745 plant chromosome Anatomy 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- -1 promoters Proteins 0.000 description 1
- 238000012175 pyrosequencing Methods 0.000 description 1
- 238000011155 quantitative monitoring Methods 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 238000009790 rate-determining step (RDS) Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 235000002020 sage Nutrition 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000003998 snake venom Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000012536 storage buffer Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000002381 testicular Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000011222 transcriptome analysis Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000009966 trimming Methods 0.000 description 1
- 208000007089 vaccinia Diseases 0.000 description 1
- 238000012418 validation experiment Methods 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
-
- 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
-
- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/682—Signal amplification
-
- 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
- C12Q1/6846—Common amplification features
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- 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
- C12Q2545/00—Reactions characterised by their quantitative nature
- C12Q2545/10—Reactions characterised by their quantitative nature the purpose being quantitative analysis
- C12Q2545/113—Reactions characterised by their quantitative nature the purpose being quantitative analysis with an external standard/control, i.e. control reaction is separated from the test/target reaction
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2021092115A JP7568581B2 (ja) | 2014-07-09 | 2021-06-01 | Rna転写産物バリアントを定量するための方法及び製品 |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP14176417.5 | 2014-07-09 | ||
| EP14176417 | 2014-07-09 | ||
| PCT/EP2015/065756 WO2016005524A1 (en) | 2014-07-09 | 2015-07-09 | Methods and products for quantifying rna transcript variants |
Related Child Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019182851A Division JP2020014478A (ja) | 2014-07-09 | 2019-10-03 | Rna転写産物バリアントを定量するための方法及び製品 |
| JP2021092115A Division JP7568581B2 (ja) | 2014-07-09 | 2021-06-01 | Rna転写産物バリアントを定量するための方法及び製品 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2017525341A JP2017525341A (ja) | 2017-09-07 |
| JP6930910B2 true JP6930910B2 (ja) | 2021-09-01 |
Family
ID=51224706
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2017501008A Active JP6930910B2 (ja) | 2014-07-09 | 2015-07-09 | Rna転写産物バリアントを定量するための方法及び製品 |
| JP2019182851A Pending JP2020014478A (ja) | 2014-07-09 | 2019-10-03 | Rna転写産物バリアントを定量するための方法及び製品 |
| JP2021092115A Active JP7568581B2 (ja) | 2014-07-09 | 2021-06-01 | Rna転写産物バリアントを定量するための方法及び製品 |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019182851A Pending JP2020014478A (ja) | 2014-07-09 | 2019-10-03 | Rna転写産物バリアントを定量するための方法及び製品 |
| JP2021092115A Active JP7568581B2 (ja) | 2014-07-09 | 2021-06-01 | Rna転写産物バリアントを定量するための方法及び製品 |
Country Status (10)
| Country | Link |
|---|---|
| US (1) | US10513726B2 (enExample) |
| EP (1) | EP3167076B1 (enExample) |
| JP (3) | JP6930910B2 (enExample) |
| KR (1) | KR102555447B1 (enExample) |
| CN (1) | CN106471134B (enExample) |
| AU (1) | AU2015286672B2 (enExample) |
| CA (1) | CA2954495C (enExample) |
| DK (1) | DK3167076T3 (enExample) |
| LT (1) | LT3167076T (enExample) |
| WO (1) | WO2016005524A1 (enExample) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2012307282B2 (en) | 2011-09-16 | 2018-03-15 | Lexogen Gmbh | Nucleic acid transcription method |
| AU2015286672B2 (en) | 2014-07-09 | 2021-12-02 | Lexogen Gmbh | Methods and products for quantifying RNA transcript variants |
| CN106771194B (zh) * | 2017-01-18 | 2018-07-20 | 青岛大学 | 一种检测病原微生物的方法 |
| US10828330B2 (en) * | 2017-02-22 | 2020-11-10 | IO Bioscience, Inc. | Nucleic acid constructs comprising gene editing multi-sites and uses thereof |
| CA3054307A1 (en) | 2017-02-22 | 2018-08-30 | Io Biosciences, Inc. | Nucleic acid constructs comprising gene editing multi-sites and uses thereof |
| SE541799C2 (en) * | 2018-04-11 | 2019-12-17 | David Yudovich | Determination of frequency distribution of nucleotide sequence variants |
| EP3781213A4 (en) | 2018-04-17 | 2022-10-19 | The Trustees of the University of Pennsylvania | TRANS-SPLICE MOLECULES |
| CN116219037A (zh) * | 2018-05-02 | 2023-06-06 | 宁夏农林科学院动物科学研究所 | 一种基于基因分型鉴定滩羊与非滩羊的方法 |
| EP3666904A1 (en) | 2018-12-14 | 2020-06-17 | Lexogen GmbH | Nucleic acid amplification and identification method |
| EP3901284A1 (en) * | 2020-04-20 | 2021-10-27 | BMBK Invest BV | Construct, kit and method for sample tracking |
| KR102690574B1 (ko) | 2020-06-26 | 2024-07-31 | 숙명여자대학교산학협력단 | 비타민 b1의 보호 및 전달을 위한 장용성 조성물 |
| KR102690573B1 (ko) | 2020-06-26 | 2024-07-31 | 숙명여자대학교산학협력단 | 사이클로덱스트린을 이용한 경구투여용 복합 조성물 및 이의 제조방법 |
| WO2023086441A1 (en) * | 2021-11-12 | 2023-05-19 | Regents Of The University Of Minnesota | Compositions and methods for transcriptional activation |
| EP4438741A4 (en) * | 2021-11-26 | 2025-08-06 | Daan Gene Co Ltd | HIGH-THROUGHPUT SEQUENCING METHOD BASED ON THE INTERNAL REFERENCE OF A KNOWN MARKER |
| CA3252043A1 (en) * | 2022-05-13 | 2023-11-16 | Ascidian Therapeutics, Inc. | ABCA4 TRANS-SPLICING MOLECULES |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2593343B1 (fr) | 1986-01-20 | 1988-03-25 | Thomson Csf | Matrice d'elements photosensibles et son procede de fabrication, procede de lecture associe, et application de cette matrice a la prise de vue d'images |
| WO2002072889A2 (en) * | 2001-01-12 | 2002-09-19 | Applera Corporation | Methods and compositions for microarray control |
| EP1386009A4 (en) | 2001-05-07 | 2005-04-27 | Amersham Biosciences Corp | CONSTRUCTION OF ARTIFICIAL GENES FOR USE AS CONTROLS IN GENE EXPRESSION ANALYSIS SYSTEMS |
| US20040009512A1 (en) | 2002-05-02 | 2004-01-15 | Manuel Ares | Arrays for detection of products of mRNA splicing |
| EP1730162A4 (en) * | 2004-03-11 | 2008-12-10 | Us Gov Health & Human Serv | ARTIFICIAL MUTATIONAL CHECKS FOR DIAGNOSTIC TESTS |
| EP2333104A1 (en) * | 2009-12-11 | 2011-06-15 | Lexogen GmbH | RNA analytics method |
| AU2015286672B2 (en) | 2014-07-09 | 2021-12-02 | Lexogen Gmbh | Methods and products for quantifying RNA transcript variants |
-
2015
- 2015-07-09 AU AU2015286672A patent/AU2015286672B2/en active Active
- 2015-07-09 US US15/323,951 patent/US10513726B2/en active Active
- 2015-07-09 JP JP2017501008A patent/JP6930910B2/ja active Active
- 2015-07-09 DK DK15736490.2T patent/DK3167076T3/da active
- 2015-07-09 EP EP15736490.2A patent/EP3167076B1/en active Active
- 2015-07-09 CA CA2954495A patent/CA2954495C/en active Active
- 2015-07-09 KR KR1020177002619A patent/KR102555447B1/ko active Active
- 2015-07-09 LT LTEPPCT/EP2015/065756T patent/LT3167076T/lt unknown
- 2015-07-09 CN CN201580035408.5A patent/CN106471134B/zh active Active
- 2015-07-09 WO PCT/EP2015/065756 patent/WO2016005524A1/en not_active Ceased
-
2019
- 2019-10-03 JP JP2019182851A patent/JP2020014478A/ja active Pending
-
2021
- 2021-06-01 JP JP2021092115A patent/JP7568581B2/ja active Active
Also Published As
| Publication number | Publication date |
|---|---|
| KR20170028383A (ko) | 2017-03-13 |
| KR102555447B1 (ko) | 2023-07-13 |
| JP2017525341A (ja) | 2017-09-07 |
| DK3167076T3 (da) | 2022-02-21 |
| US10513726B2 (en) | 2019-12-24 |
| WO2016005524A1 (en) | 2016-01-14 |
| AU2015286672A1 (en) | 2016-12-15 |
| JP2020014478A (ja) | 2020-01-30 |
| JP2021153588A (ja) | 2021-10-07 |
| HK1231518A1 (zh) | 2017-12-22 |
| EP3167076A1 (en) | 2017-05-17 |
| JP7568581B2 (ja) | 2024-10-16 |
| CA2954495C (en) | 2024-03-05 |
| CN106471134A (zh) | 2017-03-01 |
| US20170321248A1 (en) | 2017-11-09 |
| CN106471134B (zh) | 2021-11-12 |
| LT3167076T (lt) | 2022-02-10 |
| EP3167076B1 (en) | 2021-11-17 |
| CA2954495A1 (en) | 2016-01-14 |
| AU2015286672B2 (en) | 2021-12-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7568581B2 (ja) | Rna転写産物バリアントを定量するための方法及び製品 | |
| Chen et al. | RNA editome in rhesus macaque shaped by purifying selection | |
| Pfeiffer et al. | Systematic evaluation of error rates and causes in short samples in next-generation sequencing | |
| Hiller et al. | Pre-mRNA secondary structures influence exon recognition | |
| Kozarewa et al. | Overview of target enrichment strategies | |
| Lipson et al. | Quantification of the yeast transcriptome by single-molecule sequencing | |
| Gao et al. | CIRI: an efficient and unbiased algorithm for de novo circular RNA identification | |
| Li et al. | Quality control of RNA-seq experiments | |
| DK2245187T3 (en) | Methods for accurate sequence data and modified due to localization | |
| CN118638898A (zh) | 用于靶向核酸序列富集的方法及在错误纠正的核酸测序中的应用 | |
| US20140221218A1 (en) | Methods for single-molecule analysis | |
| WO2012027446A2 (en) | Nucleic acid sequence analysis | |
| Wang et al. | Forensic nanopore sequencing of microhaplotype markers using QitanTech’s QNome | |
| Bedre et al. | New era in plant alternative splicing analysis enabled by advances in high-throughput sequencing (HTS) technologies | |
| JP5926189B2 (ja) | Rna分析方法 | |
| WO2014128453A1 (en) | Nucleic acid marker molecules for identifying and detecting cross contamination of nucleic acid samples | |
| Zheng et al. | Titration-free massively parallel pyrosequencing using trace amounts of starting material | |
| US20210102246A1 (en) | Genetic test for detecting congenital adrenal hyperplasia | |
| Scheer et al. | High-Resolution mapping of 3’extremities of RNA exosome substrates by 3’RACE-seq | |
| Nikiforova et al. | Amplification-based methods | |
| Olsen et al. | Transcriptomics: RNA-seq | |
| Shin et al. | Novel discovery of line-1 in a korean individual by a target enrichment method | |
| Jain et al. | Emerging Tools for Generating Genomics Data | |
| Olsen et al. | RNA-seq | |
| Vecoli | Next-generation sequencing technology in the genetics of cardiovascular disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20170707 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20180703 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20181002 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20190104 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20190604 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20191003 |
|
| C60 | Trial request (containing other claim documents, opposition documents) |
Free format text: JAPANESE INTERMEDIATE CODE: C60 Effective date: 20191003 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20191004 |
|
| A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20191024 |
|
| C21 | Notice of transfer of a case for reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C21 Effective date: 20191029 |
|
| A912 | Re-examination (zenchi) completed and case transferred to appeal board |
Free format text: JAPANESE INTERMEDIATE CODE: A912 Effective date: 20191227 |
|
| C211 | Notice of termination of reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C211 Effective date: 20200107 |
|
| C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20200714 |
|
| C13 | Notice of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: C13 Effective date: 20201201 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20210225 |
|
| C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20210413 |
|
| C302 | Record of communication |
Free format text: JAPANESE INTERMEDIATE CODE: C302 Effective date: 20210421 |
|
| C302 | Record of communication |
Free format text: JAPANESE INTERMEDIATE CODE: C302 Effective date: 20210527 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210601 |
|
| C23 | Notice of termination of proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C23 Effective date: 20210608 |
|
| C03 | Trial/appeal decision taken |
Free format text: JAPANESE INTERMEDIATE CODE: C03 Effective date: 20210713 |
|
| C30A | Notification sent |
Free format text: JAPANESE INTERMEDIATE CODE: C3012 Effective date: 20210713 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20210812 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 6930910 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |