DE69634696T2 - Verfahren und testsystem zur hybridisierungsanalyse unter verwendung von peptidnukleinsäure-sonden - Google Patents
Verfahren und testsystem zur hybridisierungsanalyse unter verwendung von peptidnukleinsäure-sonden Download PDFInfo
- Publication number
- DE69634696T2 DE69634696T2 DE69634696T DE69634696T DE69634696T2 DE 69634696 T2 DE69634696 T2 DE 69634696T2 DE 69634696 T DE69634696 T DE 69634696T DE 69634696 T DE69634696 T DE 69634696T DE 69634696 T2 DE69634696 T2 DE 69634696T2
- Authority
- DE
- Germany
- Prior art keywords
- pna
- dna
- probe
- nucleic acid
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims description 58
- 108091093037 Peptide nucleic acid Proteins 0.000 title description 193
- 238000009396 hybridization Methods 0.000 title description 55
- 238000004458 analytical method Methods 0.000 title description 20
- 238000012360 testing method Methods 0.000 title description 3
- 239000000523 sample Substances 0.000 claims description 289
- 150000007523 nucleic acids Chemical class 0.000 claims description 59
- 102000039446 nucleic acids Human genes 0.000 claims description 54
- 108020004707 nucleic acids Proteins 0.000 claims description 54
- 230000000295 complement effect Effects 0.000 claims description 35
- 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 28
- 238000001962 electrophoresis Methods 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 23
- 238000005251 capillar electrophoresis Methods 0.000 claims description 17
- 102000040430 polynucleotide Human genes 0.000 claims description 16
- 108091033319 polynucleotide Proteins 0.000 claims description 16
- 239000002157 polynucleotide Substances 0.000 claims description 16
- 229960002685 biotin Drugs 0.000 claims description 15
- 235000020958 biotin Nutrition 0.000 claims description 15
- 239000011616 biotin Substances 0.000 claims description 15
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 11
- 150000003839 salts Chemical class 0.000 claims description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims description 10
- 239000000872 buffer Substances 0.000 claims description 9
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 8
- 229920000936 Agarose Polymers 0.000 claims description 7
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 5
- -1 fluorophores Substances 0.000 claims description 5
- 229920002401 polyacrylamide Polymers 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 239000004202 carbamide Substances 0.000 claims description 3
- 229920000609 methyl cellulose Polymers 0.000 claims description 3
- 239000001923 methylcellulose Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 230000006978 adaptation Effects 0.000 claims 1
- 239000003960 organic solvent Substances 0.000 claims 1
- 108020004414 DNA Proteins 0.000 description 116
- 238000000926 separation method Methods 0.000 description 71
- 239000000499 gel Substances 0.000 description 47
- 108090000623 proteins and genes Proteins 0.000 description 46
- 238000003556 assay Methods 0.000 description 40
- 239000012634 fragment Substances 0.000 description 37
- 238000001514 detection method Methods 0.000 description 36
- 239000002609 medium Substances 0.000 description 35
- 239000012528 membrane Substances 0.000 description 23
- 201000003883 Cystic fibrosis Diseases 0.000 description 22
- 239000002773 nucleotide Substances 0.000 description 22
- 125000003729 nucleotide group Chemical group 0.000 description 22
- 239000003298 DNA probe Substances 0.000 description 21
- 238000002474 experimental method Methods 0.000 description 20
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 17
- 238000002347 injection Methods 0.000 description 16
- 239000007924 injection Substances 0.000 description 16
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 14
- 230000035772 mutation Effects 0.000 description 13
- 230000037230 mobility Effects 0.000 description 12
- 102000053602 DNA Human genes 0.000 description 11
- 108020003215 DNA Probes Proteins 0.000 description 11
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 10
- OSBLTNPMIGYQGY-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;boric acid Chemical compound OB(O)O.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O OSBLTNPMIGYQGY-UHFFFAOYSA-N 0.000 description 9
- 230000029087 digestion Effects 0.000 description 9
- 230000005684 electric field Effects 0.000 description 9
- 238000013401 experimental design Methods 0.000 description 9
- 238000011068 loading method Methods 0.000 description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000011543 agarose gel Substances 0.000 description 7
- 238000001818 capillary gel electrophoresis Methods 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000005370 electroosmosis Methods 0.000 description 7
- 238000001499 laser induced fluorescence spectroscopy Methods 0.000 description 7
- 238000002844 melting Methods 0.000 description 7
- 230000008018 melting Effects 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 7
- 241000894007 species Species 0.000 description 7
- 108020004635 Complementary DNA Proteins 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000007983 Tris buffer Substances 0.000 description 6
- 238000000137 annealing Methods 0.000 description 6
- 108091008146 restriction endonucleases Proteins 0.000 description 6
- 238000007873 sieving Methods 0.000 description 6
- 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 5
- 239000003391 RNA probe Substances 0.000 description 5
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 5
- 238000002105 Southern blotting Methods 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000013508 migration Methods 0.000 description 5
- 230000005012 migration Effects 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- 229910052710 silicon Inorganic materials 0.000 description 5
- 239000010703 silicon Substances 0.000 description 5
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 5
- 239000002699 waste material Substances 0.000 description 5
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 4
- 239000004677 Nylon Substances 0.000 description 4
- 108020004518 RNA Probes Proteins 0.000 description 4
- 108020004682 Single-Stranded DNA Proteins 0.000 description 4
- 239000012491 analyte Substances 0.000 description 4
- 238000010804 cDNA synthesis Methods 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 239000000470 constituent Substances 0.000 description 4
- 239000000975 dye Substances 0.000 description 4
- 238000004949 mass spectrometry Methods 0.000 description 4
- 229920001778 nylon Polymers 0.000 description 4
- 230000005855 radiation Effects 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 3
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 239000002853 nucleic acid probe Substances 0.000 description 3
- 239000002751 oligonucleotide probe Substances 0.000 description 3
- 229920002120 photoresistant polymer Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- ZTDTUZROMGLNBA-UHFFFAOYSA-N 2-(4-nitrophenyl)-1h-pyrazol-3-one Chemical compound C1=CC([N+](=O)[O-])=CC=C1N1C(=O)C=CN1 ZTDTUZROMGLNBA-UHFFFAOYSA-N 0.000 description 2
- RUVRGYVESPRHSZ-UHFFFAOYSA-N 2-[2-(2-azaniumylethoxy)ethoxy]acetate Chemical group NCCOCCOCC(O)=O RUVRGYVESPRHSZ-UHFFFAOYSA-N 0.000 description 2
- 229920000663 Hydroxyethyl cellulose Polymers 0.000 description 2
- 239000004354 Hydroxyethyl cellulose Substances 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 239000004952 Polyamide Substances 0.000 description 2
- 239000013614 RNA sample Substances 0.000 description 2
- 238000001069 Raman spectroscopy Methods 0.000 description 2
- 239000008051 TBE buffer Substances 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N argon Substances [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 229910052804 chromium Inorganic materials 0.000 description 2
- 239000011651 chromium Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000005518 electrochemistry Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 238000001668 nucleic acid synthesis Methods 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 229920002647 polyamide Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 125000006239 protecting group Chemical group 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 125000006850 spacer group Chemical group 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- QTWJRLJHJPIABL-UHFFFAOYSA-N 2-methylphenol;3-methylphenol;4-methylphenol Chemical compound CC1=CC=C(O)C=C1.CC1=CC=CC(O)=C1.CC1=CC=CC=C1O QTWJRLJHJPIABL-UHFFFAOYSA-N 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 206010052656 Cystic fibrosis carrier Diseases 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 239000004831 Hot glue Substances 0.000 description 1
- 241000511976 Hoya Species 0.000 description 1
- 229910017855 NH 4 F Inorganic materials 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000003518 caustics Substances 0.000 description 1
- 239000012501 chromatography medium Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 229930003836 cresol Natural products 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 238000011157 data evaluation Methods 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000005350 fused silica glass Substances 0.000 description 1
- 230000009395 genetic defect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 238000002218 isotachophoresis Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000155 melt Substances 0.000 description 1
- 210000004088 microvessel Anatomy 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
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 230000036963 noncompetitive effect Effects 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000033458 reproduction Effects 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- SCPYDCQAZCOKTP-UHFFFAOYSA-N silanol Chemical compound [SiH3]O SCPYDCQAZCOKTP-UHFFFAOYSA-N 0.000 description 1
- 239000013464 silicone adhesive Substances 0.000 description 1
- 238000004513 sizing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 238000007693 zone electrophoresis Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/44721—Arrangements for investigating the separated zones, e.g. localising zones by optical means
- G01N27/44726—Arrangements for investigating the separated zones, e.g. localising zones by optical means using specific dyes, markers or binding molecules
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Physics & Mathematics (AREA)
- Electrochemistry (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US495395P | 1995-10-06 | 1995-10-06 | |
| US4953 | 1995-10-06 | ||
| PCT/US1996/015918 WO1997012995A1 (en) | 1995-10-06 | 1996-10-04 | Methods and kit for hybridization analysis using peptide nucleic acid probes |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| DE69634696D1 DE69634696D1 (de) | 2005-06-09 |
| DE69634696T2 true DE69634696T2 (de) | 2006-01-19 |
Family
ID=21713369
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE69634696T Expired - Lifetime DE69634696T2 (de) | 1995-10-06 | 1996-10-04 | Verfahren und testsystem zur hybridisierungsanalyse unter verwendung von peptidnukleinsäure-sonden |
Country Status (5)
| Country | Link |
|---|---|
| US (2) | US20020012902A1 (https=) |
| EP (1) | EP0854937B9 (https=) |
| JP (1) | JPH11512617A (https=) |
| DE (1) | DE69634696T2 (https=) |
| WO (1) | WO1997012995A1 (https=) |
Families Citing this family (46)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998010097A2 (en) * | 1996-09-03 | 1998-03-12 | Pioneer Hi-Bred International, Inc. | Method and apparatus for single molecule two color fluorescent detection and molecular weight and concentration determination |
| JP4110223B2 (ja) * | 1997-02-27 | 2008-07-02 | インジェネウス・インコーポレーテッド | Pnaプローブを用いる、溶液中でのヌクレオチドアッセイ法 |
| US5846729A (en) * | 1997-02-27 | 1998-12-08 | Lorne Park Research, Inc. | Assaying nucleotides in solution using a fluorescent intensity quenching effect |
| US6046004A (en) * | 1997-02-27 | 2000-04-04 | Lorne Park Research, Inc. | Solution hybridization of nucleic acids with antisense probes having modified backbones |
| US6251591B1 (en) * | 1997-02-27 | 2001-06-26 | Lorne Park Research, Inc. | Quantitative method for detecting nucleotide concentration |
| US6060242A (en) * | 1997-02-27 | 2000-05-09 | Lorne Park Research, Inc. | PNA diagnostic methods |
| DE69834038D1 (de) | 1997-07-01 | 2006-05-18 | Isis Pharmaceutical Inc | Zusammensetzungen und verfahren zur verabreichung von oligonukleotiden über die speiseröhre |
| US20030165953A1 (en) * | 1997-07-09 | 2003-09-04 | Isao Karube | Method for detecting DNA with probe PNA |
| US6255050B1 (en) | 1998-05-22 | 2001-07-03 | Lorne Park Research, Inc. | Dynamic hybridization system |
| DE69938400T2 (de) * | 1998-12-08 | 2009-04-23 | Boston Probes, Inc., Bedford | Verfahren, Kits und Zusammensetzungen zur Identifizierung von elektrostatisch an Matrizen gebundenen Nukleinsäuren |
| WO2000034521A1 (en) * | 1998-12-08 | 2000-06-15 | Boston Probes, Inc. | Methods, kits and compositions for the identification of nucleic acids electrostatically bound to matrices |
| US6361671B1 (en) | 1999-01-11 | 2002-03-26 | The Regents Of The University Of California | Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels |
| JP2002540401A (ja) * | 1999-03-19 | 2002-11-26 | ザ ガヴァナーズ オブ ザ ユニヴァーシティー オブ アルバータ | 生物学的及びその他の試料の自動化2次元分析 |
| US6645733B1 (en) | 1999-06-25 | 2003-11-11 | Ingeneus Corporation | Fluorescent intensity method for assaying binding between proteins or peptides |
| US6312956B1 (en) | 1999-10-01 | 2001-11-06 | Vanderbilt University | Nuclear targeted peptide nucleic acid oligomer |
| US7052844B2 (en) * | 1999-12-21 | 2006-05-30 | Ingeneus, Inc. | Purification of DS-DNA using heteropolymeric capture probes and a triplex, quadruplex or homologous duplex binding mechanism |
| EP1392853B1 (en) | 2000-08-30 | 2006-04-26 | Avi Biopharma, Inc. | Method for analysis of oligonucleotide analogs |
| EP1320745A4 (en) * | 2000-09-01 | 2008-10-01 | Applera Corp | SYSTEM AND METHOD FOR TEMPERATURE GRADIENT CAPILLARY ELECTROPHORESIS |
| AU2002214569A1 (en) * | 2000-10-11 | 2002-04-22 | Spectrumedix Corporation | System and method for determining the presence of methylated cytosines in polynucleotides |
| EP1432826B1 (en) | 2001-09-24 | 2012-12-12 | Life Technologies Corporation | Methods, kits and compositions pertaining to the suppression of detectable probe binding to randomly distributed repeat sequences in genomic nucleic acid |
| WO2003033743A1 (en) * | 2001-10-18 | 2003-04-24 | Spectrumedix Corporation | System and method for temperature gradient capillary electrophoresis |
| AU2002348099A1 (en) * | 2001-10-26 | 2003-05-06 | Sequenom Inc. | Resin dispensing device |
| GB0205455D0 (en) | 2002-03-07 | 2002-04-24 | Molecular Sensing Plc | Nucleic acid probes, their synthesis and use |
| WO2003080857A2 (en) * | 2002-03-21 | 2003-10-02 | Boston Probes,Inc. | Pna oligomers, oligomers sets, methods and kits pertaining to the detection of bacillus anthracis |
| US6872530B2 (en) * | 2002-04-24 | 2005-03-29 | Spectrumedix, Llc | Method for determining the presence of DNA variants using peptide nucleic acid probes |
| WO2003104383A1 (en) * | 2002-06-07 | 2003-12-18 | Millipore Corporation | Kit and process for microbiological for on-site examination of a liquid sample |
| US20040072217A1 (en) * | 2002-06-17 | 2004-04-15 | Affymetrix, Inc. | Methods of analysis of linkage disequilibrium |
| WO2004007690A2 (en) * | 2002-07-16 | 2004-01-22 | Spectrumedix Llc | Method and system for comparative genomics for organisms using temperature gradient electrophoresis |
| US7049073B2 (en) | 2002-10-30 | 2006-05-23 | The University Of Chicago | Double stranded nucleic acid biochips |
| JP4361057B2 (ja) * | 2003-04-30 | 2009-11-11 | モイル,ウィリアム,アール | 生体分子検出および細胞分類用センサ |
| US7303879B2 (en) * | 2003-07-31 | 2007-12-04 | Applera Corporation | Determination of SNP allelic frequencies using temperature gradient electrophoresis |
| US7897747B2 (en) * | 2006-05-25 | 2011-03-01 | The Board Of Trustees Of The Leland Stanford Junior University | Method to produce single stranded DNA of defined length and sequence and DNA probes produced thereby |
| EP1995328A1 (en) * | 2007-05-22 | 2008-11-26 | Koninklijke Philips Electronics N.V. | Method of detecting probe-target binding |
| US20090215050A1 (en) * | 2008-02-22 | 2009-08-27 | Robert Delmar Jenison | Systems and methods for point-of-care amplification and detection of polynucleotides |
| EP2315015A4 (en) * | 2008-07-22 | 2012-02-08 | Arkray Inc | DEVICE FOR ANALYSIS BY A CAPILLARY ELECTROPHORESIS METHOD |
| JP5395076B2 (ja) * | 2008-07-22 | 2014-01-22 | アークレイ株式会社 | キャピラリー電気泳動法による分析装置および分析方法 |
| WO2010141131A1 (en) | 2009-06-04 | 2010-12-09 | Lockheed Martin Corporation | Multiple-sample microfluidic chip for dna analysis |
| GB2497501A (en) | 2010-10-15 | 2013-06-12 | Lockheed Corp | Micro fluidic optic design |
| US9322054B2 (en) | 2012-02-22 | 2016-04-26 | Lockheed Martin Corporation | Microfluidic cartridge |
| CN109923417B (zh) | 2016-09-08 | 2021-06-08 | 希米斯保健股份有限公司 | 诊断系统和方法 |
| US10349589B2 (en) | 2016-09-08 | 2019-07-16 | Hemex Health, Inc. | Diagnostics systems and methods |
| US10538808B2 (en) | 2017-05-26 | 2020-01-21 | Vibrant Holdings, Llc | Photoactive compounds and methods for biomolecule detection and sequencing |
| WO2019060633A2 (en) * | 2017-09-22 | 2019-03-28 | Prozyme, Inc. | USE OF POLYAMINES AS RESOLUTION AGENTS FOR CAPILLARY ELECTROPHORESIS OF GLYCANES MARKETED USING GELS |
| EP3790985B1 (en) | 2018-05-09 | 2026-04-15 | Vibrant Holdings, LLC | Methods of synthesizing a polynucleotide array using photactivated agents |
| WO2020264182A1 (en) | 2019-06-25 | 2020-12-30 | Hemex Health, Inc. | Diagnostics systems and methods |
| CN112326762B (zh) * | 2020-10-29 | 2023-03-21 | 常州大学 | 一种毛细管电泳检测dna四面体的方法 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK51092D0 (da) * | 1991-05-24 | 1992-04-15 | Ole Buchardt | Oligonucleotid-analoge betegnet pna, monomere synthoner og fremgangsmaade til fremstilling deraf samt anvendelser deraf |
| US6045995A (en) * | 1994-08-24 | 2000-04-04 | Isis Pharmaceuticals, Inc. | Capillary electrophoretic detection of nucleic acids |
| US5582705A (en) * | 1995-05-19 | 1996-12-10 | Iowa State University Research Foundation, Inc. | Multiplexed capillary electrophoresis system |
-
1996
- 1996-10-04 JP JP9514448A patent/JPH11512617A/ja not_active Ceased
- 1996-10-04 US US08/726,093 patent/US20020012902A1/en not_active Abandoned
- 1996-10-04 EP EP96934037A patent/EP0854937B9/en not_active Expired - Lifetime
- 1996-10-04 WO PCT/US1996/015918 patent/WO1997012995A1/en not_active Ceased
- 1996-10-04 DE DE69634696T patent/DE69634696T2/de not_active Expired - Lifetime
-
2003
- 2003-08-07 US US10/636,343 patent/US20050053944A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| WO1997012995A1 (en) | 1997-04-10 |
| DE69634696D1 (de) | 2005-06-09 |
| US20050053944A1 (en) | 2005-03-10 |
| EP0854937B9 (en) | 2006-06-28 |
| JPH11512617A (ja) | 1999-11-02 |
| EP0854937A1 (en) | 1998-07-29 |
| EP0854937B1 (en) | 2005-05-04 |
| US20020012902A1 (en) | 2002-01-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE69634696T2 (de) | Verfahren und testsystem zur hybridisierungsanalyse unter verwendung von peptidnukleinsäure-sonden | |
| DE68909514T2 (de) | Verfahren zur gleichzeitigen Bestimmung von DNS-Sequenzvariationen von zahlreichen Stellen und ein Satz dafür. | |
| DE60222427T2 (de) | Puffer-arrays zum nachweis von biomolekülen anhand ihres isoelektrischen punktes | |
| DE69233458T2 (de) | Nukleinsäuretypisierung durch polymeraseverlängerung von oligonukleotiden unter verwendung von terminator-mischungen | |
| DE69713599T2 (de) | Verfahren zur bestimmung von nukleinsäure-sequenzen und diagnostische anwendungen derselben | |
| DE69617274T2 (de) | Verfahren und vorrichtung für diagnostischen dns-test | |
| DE69314946T2 (de) | Verfahren und zusammensetzung für den nachweis von verschiedenen dns sequenzen | |
| EP0925373B1 (de) | Verfahren zum nachweis von nukleinsäuren unter ermittlung der masse | |
| WO1997012995A9 (en) | Methods and kit for hybridization analysis using peptide nucleic acid probes | |
| DE60127469T2 (de) | Potentiometrischer dna-mikroarray, verfahren zu dessen herstellung und verfahren zur nukleinsäureanalyse | |
| DE69221980T2 (de) | Verfahren und vorrichtung zur bestimmung von dns-nukleotid-sequenzen | |
| DE60208278T2 (de) | System und Verfahren zum Testen von Nukleinsäuremolekülen | |
| DE69428000T2 (de) | Analysen- und trennungsverfahren | |
| EP1381698A2 (de) | Verfahren zur analyse von nukleinsäureketten | |
| WO2003020968A2 (de) | Verfahren zur analyse von nukleinsäurekettensequenzen und der genexpression | |
| WO2002088381A2 (de) | Verfahren zur bestimmung der genexpression | |
| DE69814848T2 (de) | Untersuchung von nukleotiden in loesung mit hilfe von pna-sonden | |
| EP0486580B1 (de) | Verfahren und vorrichtung zur trennung und detektion von komponenten eines stoffgemisches durch temperaturgradienten-gelelektrophorese | |
| EP1565569A2 (de) | Verfahren zum nachweis von mutationen | |
| EP1186669B1 (de) | Verfahren zur spezifischen Bestimmung von DNA-Sequenzen mittels paralleler Amplifikation | |
| DE10259819B4 (de) | Verfahren zur PCR-Amplifikation und Detektion von Nucleotidsequenzen | |
| DE69724375T2 (de) | Verfahren zur Messung der Polynukleotidkonzentration | |
| EP1477572A2 (en) | Methods and kit for hybridization analysis using peptide nucleic acid probes | |
| EP1441035B1 (de) | Verfahren zur Erstellung von Profilen der differentialen Proteinexpression | |
| DE19957319A1 (de) | Dynamische Bestimmung von Analyten |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 8364 | No opposition during term of opposition | ||
| 8327 | Change in the person/name/address of the patent owner |
Owner name: APPLIED BIOSYSTEMS, LLC (N. D. GES. D. STAATES, US |