CN115260261A - Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye - Google Patents
Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye Download PDFInfo
- Publication number
- CN115260261A CN115260261A CN202110479298.9A CN202110479298A CN115260261A CN 115260261 A CN115260261 A CN 115260261A CN 202110479298 A CN202110479298 A CN 202110479298A CN 115260261 A CN115260261 A CN 115260261A
- Authority
- CN
- China
- Prior art keywords
- compound
- deoxynucleoside
- fluorescent dye
- solid
- organic solvent
- 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.)
- Pending
Links
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 42
- 239000007790 solid phase Substances 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 52
- 150000001875 compounds Chemical class 0.000 claims abstract description 30
- 239000003960 organic solvent Substances 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 28
- -1 diamine trifluoroacetamide derivative Chemical class 0.000 claims abstract description 16
- 150000002148 esters Chemical class 0.000 claims abstract description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229940125782 compound 2 Drugs 0.000 claims abstract description 15
- 229940126214 compound 3 Drugs 0.000 claims abstract description 15
- 229940125898 compound 5 Drugs 0.000 claims abstract description 14
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 13
- 239000000126 substance Substances 0.000 claims abstract description 10
- 150000008065 acid anhydrides Chemical class 0.000 claims abstract description 6
- 150000007530 organic bases Chemical class 0.000 claims abstract description 6
- JBWYRBLDOOOJEU-UHFFFAOYSA-N 1-[chloro-(4-methoxyphenyl)-phenylmethyl]-4-methoxybenzene Chemical compound C1=CC(OC)=CC=C1C(Cl)(C=1C=CC(OC)=CC=1)C1=CC=CC=C1 JBWYRBLDOOOJEU-UHFFFAOYSA-N 0.000 claims abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 48
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 24
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 23
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 15
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 13
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 12
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 6
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 6
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- YWQZUENVQOQEHJ-PIXDULNESA-N (e)-3-[1-[(2r,4s,5r)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-2,4-dioxopyrimidin-5-yl]prop-2-enoic acid Chemical group C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(\C=C\C(O)=O)=C1 YWQZUENVQOQEHJ-PIXDULNESA-N 0.000 claims description 5
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 4
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 4
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 3
- 239000011324 bead Substances 0.000 claims description 3
- 239000000975 dye Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 229940014800 succinic anhydride Drugs 0.000 claims description 3
- 229930024421 Adenine Natural products 0.000 claims description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 2
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 claims description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- 229960000643 adenine Drugs 0.000 claims description 2
- 125000004202 aminomethyl group Chemical group [H]N([H])C([H])([H])* 0.000 claims description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 2
- 150000008064 anhydrides Chemical class 0.000 claims description 2
- 229940104302 cytosine Drugs 0.000 claims description 2
- VANNPISTIUFMLH-UHFFFAOYSA-N glutaric anhydride Chemical compound O=C1CCCC(=O)O1 VANNPISTIUFMLH-UHFFFAOYSA-N 0.000 claims description 2
- AALHTIXHXHZNTJ-UHFFFAOYSA-N pentane-1,5-diamine;2,2,2-trifluoroacetamide Chemical compound NCCCCCN.NC(=O)C(F)(F)F AALHTIXHXHZNTJ-UHFFFAOYSA-N 0.000 claims description 2
- 229920005990 polystyrene resin Polymers 0.000 claims description 2
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 2
- 235000015497 potassium bicarbonate Nutrition 0.000 claims description 2
- 239000011736 potassium bicarbonate Substances 0.000 claims description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 2
- 235000011181 potassium carbonates Nutrition 0.000 claims description 2
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 2
- 239000001022 rhodamine dye Chemical class 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- 235000017550 sodium carbonate Nutrition 0.000 claims description 2
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical class [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 claims description 2
- 229940113082 thymine Drugs 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 description 16
- 239000002904 solvent Substances 0.000 description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 239000012074 organic phase Substances 0.000 description 8
- 238000010791 quenching Methods 0.000 description 8
- 230000000171 quenching effect Effects 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 238000004440 column chromatography Methods 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 238000001704 evaporation Methods 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 229960001701 chloroform Drugs 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000004821 distillation Methods 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 229940125904 compound 1 Drugs 0.000 description 3
- UAOMVDZJSHZZME-UHFFFAOYSA-N diisopropylamine Chemical compound CC(C)NC(C)C UAOMVDZJSHZZME-UHFFFAOYSA-N 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 239000003298 DNA probe Substances 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 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 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108020003215 DNA Probes Proteins 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 229940043279 diisopropylamine Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 150000008300 phosphoramidites Chemical class 0.000 description 1
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/073—Pyrimidine radicals with 2-deoxyribosyl as the saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
- C09K11/025—Use of particular materials as binders, particle coatings or suspension media therefor non-luminescent particle coatings or suspension media
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Materials Engineering (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention relates to a preparation method of a deoxynucleoside solid-phase carrier modified by a fluorescent dye, which comprises the following steps: dissolving deoxynucleoside into a first organic solvent, and adding 4,4' -dimethoxytriphenylchloromethane to obtain a compound 2; dissolving the compound 2 into a second organic solvent, and adding a first condensing agent and a diamine trifluoroacetamide derivative to obtain a compound 3; dissolving the compound 3 into a methanol solution, adding an alkaline substance, and reacting to obtain a compound 4; dissolving the compound 4 into a third organic solvent, and adding fluorescent dye activated ester to obtain a compound 5; dissolving the compound 5 in a fourth organic solvent, and sequentially adding acid anhydride and organic base to obtain a compound 6; and dissolving the compound 6 into a fifth organic solvent, sequentially adding a second condensing agent and the solid phase carrier, and separating the deoxynucleoside solid phase carrier modified by the fluorescent dye after reaction. The method is simple and convenient, has high yield and realizes the aim of efficient fluorescence diagnosis.
Description
Technical Field
The invention belongs to the field of biomolecule diagnosis, relates to a synthetic method of a fluorescent group in a fluorescent probe, and particularly relates to a preparation method of a deoxynucleoside solid-phase carrier modified by a fluorescent dye.
Background
The fluorescent probe is characterized in that fluorescent substances are used as indicators, the fluorescent substances generate fluorescence under the excitation of certain wavelength, and the detected substances are quantitatively or qualitatively analyzed by detecting the generated fluorescence. In the aspect of biomedical detection, compared with other detection methods, the fluorescent probe has the advantages of high sensitivity, high selectivity, quick response, good repeatability, simplicity and convenience in operation, low cost and the like, so that the fluorescent probe is widely applied to the aspects of molecular diagnosis, gene detection, antibody immunoassay and the like.
The fluorescent probe consists of three parts, namely a recognition group, a fluorescent group and a connector, wherein the recognition group determines the selectivity and specificity of the probe, the fluorescent group determines the sensitivity of the probe, and the connector part plays a role in connecting a hub in the probe. The fluorescent dye exists in the fluorescent probe in the identity of a fluorescent group, and the photophysical property of the fluorescent dye is important to the performance of the probe. At present, rhodamine, fluorescein, phthalic aldehyde and the like are mainly used as fluorescent probes for labeling or derivation, wherein the fluorescein compound occupies an important position in the field of biological research. However, the preparation method of the deoxynucleoside solid phase carrier modified by the fluorescent dye is not mature in China.
Disclosure of Invention
The application mainly solves the problem that the preparation method of the deoxynucleoside solid-phase carrier modified by the fluorescent dye is not available at present.
Based on the above problems, the present application proposes the following solutions:
a preparation method of a deoxynucleoside solid phase carrier modified by a fluorescent dye comprises the following steps:
s1, fully dissolving deoxynucleoside into a first organic solvent, adding 4,4' -dimethoxytriphenylchloromethane, fully reacting, and separating out a compound 2;
s2, fully dissolving the compound 2 in the step S1 into a second organic solvent, adding a first condensing agent and a diamine trifluoroacetamide derivative, fully reacting, and separating out a compound 3;
s3, fully dissolving the compound 3 in the step S2 into a methanol solution, adding an alkaline substance, and separating a compound 4 after full reaction;
s4, fully dissolving the compound 4 in the step S3 into a third organic solvent, adding a fluorescent dye activated ester, and separating a compound 5 after full reaction;
s5, fully dissolving the compound 5 in the step S4 into a fourth organic solvent, sequentially adding acid anhydride and organic base, and separating a compound 6 after full reaction;
and S6, fully dissolving the compound 6 in the step S5 into a fifth organic solvent, sequentially adding a second condensing agent and the solid phase carrier, and separating the deoxynucleoside solid phase carrier modified by the fluorescent dye after full reaction.
As a further improvement of the present application, in step S1, the first organic solvent may be, but is not limited to, any one of pyridine, DMF, dichloromethane, ethyl acetate, and tetrahydrofuran.
As a further improvement of the present application, in step S2, the second organic solvent may be, but is not limited to, any one of DMF, dichloromethane, chloroform, ethyl acetate, diethyl ether, and acetonitrile.
As a further improvement of the present application, in step S2, the first condensing agent may be, but is not limited to, any one of DCC, EDC, HOBt, HBTu, HATu, and PyBop.
As a further improvement of the present application, in step S2, the diamine trifluoroacetamide derivative may be, but is not limited to, any one of ethylenediamine trifluoroacetamide, propylenediamine trifluoroacetamide, butylenediamine trifluoroacetamide, pentylenediamine trifluoroacetamide, and hexylenediamine trifluoroacetamide.
As a further improvement of the present application, in step S3, the basic substance may be, but is not limited to, at least one of sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, ammonia, and methylamine.
As a further improvement of the present application, in step S4, the third organic solvent may be, but is not limited to, any one of DMF, dichloromethane, trichloromethane, ethyl acetate, diethyl ether, and acetonitrile.
As a further improvement of the present application, in step S4, the fluorescent dye activated ester may be, but is not limited to, any one of FAM activated ester, HEX activated ester, TET activated ester, cyanine dye activated ester, and rhodamine dye activated ester.
As a further improvement of the present application, in step S4, the temperature of the reaction is 60 ℃ to 90 ℃.
As a further improvement of the present application, in step S5, the fourth organic solvent may be, but is not limited to, any one of DMF, dichloromethane, trichloromethane, ethyl acetate, diethyl ether, and acetonitrile.
As a further improvement of the present application, in step S5, the organic base may be, but is not limited to, any one of triethylamine, pyridine, diisopropyl amine, and diisopropyl ethylamine.
As a further improvement of the present application, in step S5, the acid anhydride may be, but is not limited to, any one of succinic anhydride, glutaric anhydride, and diethanol anhydride.
As a further improvement of the present application, in step S6, the fifth organic solvent may be, but is not limited to, any one of DMF, dichloromethane, trichloromethane, ethyl acetate, diethyl ether, and acetonitrile.
As a further improvement of the present application, in step S6, the second condensing agent may be, but is not limited to, any one of DCC, EDC, HOBt, HBTu, HATu, and PyBop.
As a further improvement of the present application, in step S6, the solid phase carrier may be, but not limited to, any one of controlled glass bead powder CPG and aminomethyl polystyrene resin.
As a further improvement of the present application, the deoxynucleotide may be, but is not limited to, any one of adenine deoxynucleotide, thymine deoxynucleotide, cytosine deoxynucleotide and guanine deoxynucleotide.
As a further improvement herein, the deoxynucleoside may be, but is not limited to, 5- (2-carboxyvinyl) -2' -deoxyuridine.
The method has the advantages that the method for preparing the deoxynucleoside solid-phase carrier modified by the fluorescent dye is simple and convenient, the yield is high, and the purpose of efficient fluorescence diagnosis is achieved.
Drawings
FIG. 1 shows the synthetic reaction process of the deoxynucleoside solid-phase carrier modified by fluorescent dye.
Detailed Description
In order to make the objects, technical solutions and advantages of the present application more clear, the following description of the present application will be made in detail and completely with reference to the specific embodiments and the accompanying drawings. It should be understood that the described embodiments are only a few embodiments of the present application, not all embodiments, and are not intended to limit the scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
The application provides a preparation method of a deoxynucleoside solid-phase carrier modified by a fluorescent dye, which comprises the following steps:
s1, fully dissolving deoxynucleoside into a first organic solvent, adding 4,4' -dimethoxytriphenylchloromethane, fully reacting, and separating out a compound 2; preferably, the reaction is carried out at room temperature, the separation step comprises the steps of carrying out first-step decompression and first-step extraction in sequence after full reaction, collecting a first organic phase, removing the solvent, and carrying out first-step purification on the residue to obtain a compound 2; further, the extract used in the first extraction step is a mixed solution of water and dichloromethane, the solvent removal process in this step is reduced pressure distillation, and the first purification step may be, but not limited to, chromatography, column chromatography, etc.
S2, fully dissolving the compound 2 in the step S1 into a second organic solvent, adding a first condensing agent and a diamine trifluoroacetamide derivative, fully reacting, and separating out a compound 3; preferably, the reaction is carried out at room temperature, the separation step comprises the steps of carrying out first-step quenching and second-step extraction in sequence after full reaction, collecting a second organic phase, removing the solvent, and carrying out second-step purification on the residue to obtain a compound 3; furthermore, the solution used in the first quenching step is water, the extract used in the second extraction step is ethyl acetate, the solvent removal process in this step is reduced pressure distillation, and the second purification step can be, but not limited to, chromatography, column chromatography, etc.
S3, fully dissolving the compound 3 in the step S2 into a methanol solution, adding an alkaline substance, and separating a compound 4 after full reaction; preferably, the reaction is carried out at room temperature, and the separation step is to fully react and then concentrate and dry.
S4, fully dissolving the compound 4 in the step S3 into a third organic solvent, adding a fluorescent dye activated ester, and separating a compound 5 after full reaction; preferably, the reaction is carried out under heating conditions, the separation step is to remove the solvent after the reaction is completed, dichloromethane is added to the residue, a third organic phase is collected after washing with water for three times, concentration is carried out, and finally, the compound 5 is obtained after the third purification step; further, the solvent removal process in this step is reduced pressure distillation, and the purification process in the third step may be, but not limited to, chromatography, column chromatography, etc.
S5, fully dissolving the compound 5 in the step S4 into a fourth organic solvent, sequentially adding acid anhydride and organic base, fully reacting, and separating out a compound 6; preferably, the reaction is carried out at room temperature, and the separation step comprises the steps of sequentially carrying out the second quenching step and the third extraction step, collecting a fourth organic phase, and removing the solvent to obtain a compound 6; further, the liquid used in the second quenching step is water, and the solvent removal process is distillation under reduced pressure.
S6, fully dissolving the compound 6 in the step S5 into a fifth organic solvent, sequentially adding a second condensing agent and a solid phase carrier, and separating the deoxynucleoside solid phase carrier modified by the fluorescent dye after full reaction; preferably, the reaction is carried out at room temperature for 10h, and the separation step is filtration and solid collection.
Purpose of quenching reaction in the above step: in organic chemical reactions, a reaction substrate is in excess, and when the chemical reaction proceeds to a certain extent and the target product is obtained, the reaction substrate in excess is continued to react further to form a by-product or to affect the post-treatment operation, so that it needs to be quenched. Quenching is to react with another compound which is easier to react with the excessive substance, and the compound is removed from the system by reducing the reaction rate or destroying the reaction system so as to stop the reaction, thereby achieving the purposes of reducing the generation of byproducts and ensuring the safety and convenience of the post-treatment of the experiment.
In the present application, since 5- (2-carboxyvinyl) -2 '-deoxyuridine is one of deoxynucleosides, 5- (2-carboxyvinyl) -2' -deoxyuridine is described as an example, and specific examples are as follows:
a method for preparing a deoxynucleoside solid-phase carrier modified by a fluorescent dye is disclosed, wherein the overall process of the synthesis reaction is shown in figure 1, and the method comprises the following steps:
s1, 40g of compound 1 is placed in a dry flask, said compound 1 being: 5- (2-carboxyvinyl) -2 '-deoxyuridine, adding 300ml pyridine into a flask for dissolving, then adding 45 g 4,4' -dimethoxytriphenylchloromethane for reacting at room temperature till completion, evaporating pyridine under reduced pressure, then extracting with water and 300ml dichloromethane, collecting an organic phase, evaporating solvent under reduced pressure, and purifying residue by column chromatography to obtain a compound 2, wherein the compound 2 is 52 g pure white solid, the compound 1 is subjected to a reaction a to obtain a compound 2, and the reaction a is that
Nuclear magnetic detection HNMR on compound 2 was:1H NMR(DMSO)δ2.16-2.25(m,1H),2.30-2.36(m,1H),3.16(dd,1H),3.24(dd,1H),3.72(dd,6H),3.88-3.92(m,1H),4.21-4.26(m,1H),5.29(d,1H),6.17(dd,1H),6.78-6.87(m,5H),7.17-7.38(m,10H),8.03(s,1H),11.67(s,1H),12.1(w,1H)。
s2, placing 52 g of compound 2 in the step S1 into a dry flask, adding 300ml of DMF for dissolving, sequentially adding a condensing agent 27 g of DCC, 18 g of HOBt, 24ml of triethylamine and 22 g of 1, 6-hexanediamine trifluoroacetamide, reacting at room temperature till the reaction is complete, adding 800 ml of water for quenching, extracting and layering with 800 ml of ethyl acetate, collecting an organic phase, evaporating under reduced pressure to remove the solvent, purifying residues by using column chromatography to obtain 50 g of white solid compound 3, reacting the compound 2 with a reaction b to obtain a compound 3, and reacting the reaction b with the reaction b to obtain the compound 3
The nmr measurements performed on compound 3 were:1H NMR(DMSO)δ1.23-.128(m,4H),1.38-1.48(m,4H),2.14-2.20(m,1H),2.30-2.36(m,1H),2.99-3.03(m,1H),3.10-3.19(m,5H),3.72(dd,6H),3.88-3.92(m,1H),4.21-4.26(m,1H),5.29(d,1H),6.17(t,1H),6.85-6.91(m,4H),7.05-7-4(m,11H),7.94(s,1H),8.01(t,1H),9.39(t,1H),11.60(s,1H)。
s3, placing 50 g of the compound 3 in the step S2 into a dry flask, adding 100 ml of methanol for dissolving, then adding 100 ml of concentrated ammonia water, reacting at room temperature until the reaction is complete, then concentrating until the reaction is dry to obtain a compound 4, directly using the product in the next reaction, obtaining the compound 4 from the compound 3 through a reaction c, wherein the reaction c is
S4, putting 44 g of the compound 4 in the step S3 into a flask, adding 400 ml of DMF for dissolving, then adding 40g of FAM-NHS, heating to 80 ℃ for overnight reaction, evaporating the DMF solvent under reduced pressure after the reaction is finished, adding 500 ml of dichloromethane for dissolving the residue, washing for 3 times, collecting an organic phase, concentrating, purifying the residue by using a column chromatography column to obtain 40g of a compound 5, obtaining the compound 5 from the compound 4 through a reaction d, wherein the reaction d is
The nmr measurements performed on compound 5 were:1H NMR(DMSO)δ1.23-.128(m,4H),1.32(s,18H),1.38-1.48(m,4H),2.14-2.20(m,1H),2.30-2.36(m,1H),2.99-3.03(m,1H),3.10-3.19(m,5H),3.72(dd,6H),3.88-3.92(m,1H),4.21-4.26(m,1H),5.29(d,1H),6.17(t,1H),6.8(dd,4H),6.85-6.91(m,4H),7.05-7.4(m,14H),7.94(s,1H),8.01(t,1H),8.40(d,1H),8.73(s,1H),9.39(t,1H),11.60(s,1H)。
s5, placing 40g of the compound 5 in the step S4 into a dry flask, adding 400 ml of dichloromethane for dissolving, sequentially adding 4.9 g of succinic anhydride and 9 ml of triethylamine, reacting at room temperature till the reaction is complete, adding 200 ml of water for quenching, extracting and layering, collecting an organic phase, evaporating under reduced pressure to remove the solvent to obtain a compound 6, directly using the product in the next step, obtaining the compound 6 from the compound 5 through a reaction e, wherein the reaction e is that
S6, placing 10 g of the compound 6 in the step S5 into a dry flask, adding dichloromethane to dissolve the compound, then sequentially adding 8 g of DCC,5 g of HOBt,10 ml of triethylamine and controlled glass bead powder CPG, stirring at room temperature to react for 10 hours, filtering, collecting solids to obtain a compound 7, namely FAM-dT-CPG, and obtaining the compound 6 from the compound 6 through a reaction f, wherein the reaction f is as follows:
in summary, the application provides a preparation method of a deoxynucleoside solid-phase carrier modified by a fluorescent dye, the method is simple and convenient, the yield is high, and the purpose of efficient fluorescence diagnosis is realized. And FAM is a common fluorescein dye, the excitation wavelength of which is 498nm, the emission wavelength of which is 525nm, and is commonly used in fluorescent probes. FAM can also be modified on deoxynucleoside dT to form FAM-dT phosphoramidite monomer, and any site of the probe can be marked in the synthesis of DNA probe, and FAM-dT-solid phase carrier can also be loaded on solid phase carrier to form FAM-dT-solid phase carrier, such as: FAM-dT-CPG (PS) for 3' end modification of DNA probes. The invention particularly introduces a method for synthesizing a solid phase carrier of FAM-dT-CPG (PS).
Although the description is given in terms of embodiments, not every embodiment includes only a single embodiment, and such description is for clarity only, and those skilled in the art will recognize that the embodiments described herein may be combined as a whole to form other embodiments as would be understood by those skilled in the art.
The above-listed detailed description is only a specific description of a possible embodiment of the present invention, and they are not intended to limit the scope of the present invention, and equivalent embodiments or modifications made without departing from the technical spirit of the present invention should be included in the scope of the present invention.
Claims (17)
1. A preparation method of a deoxynucleoside solid phase carrier modified by a fluorescent dye is characterized by comprising the following steps:
s1, fully dissolving deoxynucleoside into a first organic solvent, adding 4,4' -dimethoxytriphenylchloromethane, fully reacting, and separating out a compound 2;
s2, fully dissolving the compound 2 in the step S1 into a second organic solvent, adding a first condensing agent and a diamine trifluoroacetamide derivative, fully reacting, and separating out a compound 3;
s3, fully dissolving the compound 3 in the step S2 into a methanol solution, adding an alkaline substance, and separating a compound 4 after full reaction;
s4, fully dissolving the compound 4 in the step S3 into a third organic solvent, adding a fluorescent dye activated ester, and separating a compound 5 after full reaction;
s5, fully dissolving the compound 5 in the step S4 into a fourth organic solvent, sequentially adding acid anhydride and organic base, and separating a compound 6 after full reaction;
and S6, fully dissolving the compound 6 in the step S5 into a fifth organic solvent, sequentially adding a second condensing agent and the solid phase carrier, and separating the deoxynucleoside solid phase carrier modified by the fluorescent dye after full reaction.
2. The method of preparing the solid phase support of a deoxynucleoside modified with a fluorescent dye according to claim 1, wherein the first organic solvent is any one of pyridine, DMF, dichloromethane, ethyl acetate, and tetrahydrofuran in step S1.
3. The method of claim 1, wherein in step S2, the second organic solvent is any one of DMF, dichloromethane, chloroform, ethyl acetate, diethyl ether, and acetonitrile.
4. The method of preparing a deoxynucleoside solid support modified with a fluorescent dye according to claim 1, wherein the first condensing agent is any one of DCC, EDC, HOBt, HBTU, HATu and PyBop in step S2.
5. The method for preparing the deoxynucleoside solid-phase carrier modified with the fluorescent dye according to claim 1, wherein in step S2, the diamine trifluoroacetamide derivative is any one of ethylenediamine trifluoroacetamide, propylenediamine trifluoroacetamide, butylenediamine trifluoroacetamide, pentylenediamine trifluoroacetamide and hexylenediamine trifluoroacetamide.
6. The method for preparing a solid phase support of a deoxynucleoside modified with a fluorescent dye according to claim 1, wherein in step S3, the basic substance is at least one of sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, ammonia water, and methylamine.
7. The method of claim 1, wherein in step S4, the third organic solvent is any one of DMF, dichloromethane, chloroform, ethyl acetate, diethyl ether, and acetonitrile.
8. The method for preparing the deoxynucleoside solid-phase carrier modified with the fluorescent dye according to claim 1, wherein in the step S4, the fluorescent dye activated ester is any one of FAM activated ester, HEX activated ester, TET activated ester, cyanine dye activated ester, and rhodamine dye activated ester.
9. The method of preparing a fluorescent dye-modified deoxynucleoside solid support according to claim 1, wherein the reaction temperature in step S4 is 60 ℃ to 90 ℃.
10. The method of claim 1, wherein in step S5, the fourth organic solvent is any one of DMF, dichloromethane, chloroform, ethyl acetate, diethyl ether, and acetonitrile.
11. The method for preparing a deoxynucleoside solid-phase carrier modified with a fluorescent dye according to claim 1, wherein in the step S5, the organic base is any one of triethylamine, pyridine, diisopropylethylamine and diisopropylethylamine.
12. The method for preparing a solid phase support of a deoxynucleoside modified with a fluorescent dye according to claim 1, wherein in the step S5, the acid anhydride is any one of succinic anhydride, glutaric anhydride and diethanol anhydride.
13. The method of claim 1, wherein in step S6, the fifth organic solvent is any one of DMF, dichloromethane, chloroform, ethyl acetate, diethyl ether, and acetonitrile.
14. The method for preparing a fluorescent dye-modified deoxynucleoside solid-phase carrier according to claim 1, wherein in step S6, the second condensing agent is any one of DCC, EDC, HOBt, HBTU, HATu and PyBop.
15. The method for preparing the deoxynucleoside solid-phase carrier modified with the fluorescent dye according to claim 1, wherein in step S6, the solid-phase carrier is any one of controlled glass bead powder CPG and aminomethyl polystyrene resin.
16. The method for preparing a deoxynucleoside solid-phase carrier modified with a fluorescent dye according to claim 1, wherein the deoxynucleotide is any one of adenine deoxynucleotide, thymine deoxynucleotide, cytosine deoxynucleotide and guanine deoxynucleotide.
17. The method of claim 1, wherein the deoxynucleoside is 5- (2-carboxyvinyl) -2' -deoxyuridine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110479298.9A CN115260261A (en) | 2021-04-30 | 2021-04-30 | Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110479298.9A CN115260261A (en) | 2021-04-30 | 2021-04-30 | Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115260261A true CN115260261A (en) | 2022-11-01 |
Family
ID=83745888
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110479298.9A Pending CN115260261A (en) | 2021-04-30 | 2021-04-30 | Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115260261A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030180757A1 (en) * | 1996-01-23 | 2003-09-25 | Affymetrix, Inc. | Nucleic acid labeling compounds |
| CN102219819A (en) * | 2011-04-19 | 2011-10-19 | 李辉 | Nucleotide monomer for improving Tm value of oligonucleotide chain and preparation method and application thereof |
| JP2013039060A (en) * | 2011-08-12 | 2013-02-28 | Saitama Univ | Linker for evolving protein with enzyme-like activity, and method for screening such protein using the linker |
| US20130066063A1 (en) * | 2011-09-09 | 2013-03-14 | John Cooke Hodges | Bicyclo[6.1.0]non-4-yne regents for chemical modification of oligonucleotides |
| US20150152496A1 (en) * | 2012-08-30 | 2015-06-04 | Kabushiki Kaisha Dnaform | Method for analyzing target nucleic acid, kit, and analyzer |
| CN105348343A (en) * | 2015-11-25 | 2016-02-24 | 北京大学 | Chromophore-modified deoxynucleoside phosphoramidite monomer compound, preparation method therefor and application thereof |
| CN109715644A (en) * | 2016-09-15 | 2019-05-03 | 日本电气方案创新株式会社 | Nucleoside derivates or its salt, the synthetic agent of polynucleotides, the manufacturing method of polynucleotides, polynucleotides and the manufacturing method for combining nucleic acid molecules |
| CN109715807A (en) * | 2016-09-15 | 2019-05-03 | 日本电气方案创新株式会社 | Secretory immunoglobulin A (sIgA) combines nucleic acid molecules, sIgA analysis sensor and sIgA analysis method |
| CN111018930A (en) * | 2019-12-06 | 2020-04-17 | 深圳大学 | TPE-2' -deoxynucleoside, fluorescent gel, and preparation methods and applications thereof |
-
2021
- 2021-04-30 CN CN202110479298.9A patent/CN115260261A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030180757A1 (en) * | 1996-01-23 | 2003-09-25 | Affymetrix, Inc. | Nucleic acid labeling compounds |
| CN102219819A (en) * | 2011-04-19 | 2011-10-19 | 李辉 | Nucleotide monomer for improving Tm value of oligonucleotide chain and preparation method and application thereof |
| JP2013039060A (en) * | 2011-08-12 | 2013-02-28 | Saitama Univ | Linker for evolving protein with enzyme-like activity, and method for screening such protein using the linker |
| US20130066063A1 (en) * | 2011-09-09 | 2013-03-14 | John Cooke Hodges | Bicyclo[6.1.0]non-4-yne regents for chemical modification of oligonucleotides |
| US20150152496A1 (en) * | 2012-08-30 | 2015-06-04 | Kabushiki Kaisha Dnaform | Method for analyzing target nucleic acid, kit, and analyzer |
| CN105348343A (en) * | 2015-11-25 | 2016-02-24 | 北京大学 | Chromophore-modified deoxynucleoside phosphoramidite monomer compound, preparation method therefor and application thereof |
| CN109715644A (en) * | 2016-09-15 | 2019-05-03 | 日本电气方案创新株式会社 | Nucleoside derivates or its salt, the synthetic agent of polynucleotides, the manufacturing method of polynucleotides, polynucleotides and the manufacturing method for combining nucleic acid molecules |
| CN109715807A (en) * | 2016-09-15 | 2019-05-03 | 日本电气方案创新株式会社 | Secretory immunoglobulin A (sIgA) combines nucleic acid molecules, sIgA analysis sensor and sIgA analysis method |
| CN111018930A (en) * | 2019-12-06 | 2020-04-17 | 深圳大学 | TPE-2' -deoxynucleoside, fluorescent gel, and preparation methods and applications thereof |
Non-Patent Citations (1)
| Title |
|---|
| KENZO FUJIMOTO，等: "Development of Photochemical DNA/RNA Manipulation Toward Its Application for Nanotechnology", 《RANSACTIONS OF THE MATERIALS R ESEARCH SOCIETY OF JAPAN》, vol. 35, no. 1, 31 December 2010 (2010-12-31), pages 85 - 89 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111205327B (en) | Preparation method of Reideciclovir | |
| DK171822B1 (en) | Modified nucleotides and methods for their preparation and detection | |
| CN114014848A (en) | RNA fluorescent probe and preparation method and application thereof | |
| WO2022166436A1 (en) | Method for preparing moxifloxacin intermediate (s, s)-2, 8-diazabicyclo [4, 3, 0] nonane | |
| CN115260261A (en) | Preparation method of deoxynucleoside solid-phase carrier modified by fluorescent dye | |
| CN107033331A (en) | A kind of fluorescence conjugated macromolecule of side-chain amino group protonation, preparation method and applications | |
| CN114685485A (en) | Synthesis method of pyridylimidazole thiopropionic acid anti-gout compound | |
| CN114634497B (en) | Cysteine/homocysteine response AIE fluorescent probe and preparation method and application thereof | |
| CN114539460B (en) | Synthesis method of macromolecule supported asymmetric catalyst | |
| CN1940563A (en) | Fluorescent marking reagent | |
| CN100460416C (en) | Method for preparing adenosine | |
| CN113372341A (en) | Synthesis method and device of dihydroisoquinoline trifluoroacetate derivative | |
| CN113336701A (en) | Nitric oxide two-photon lipid droplet locking fluorescent probe, preparation method thereof and application thereof in detecting neuroinflammation | |
| CN111205210A (en) | Synthetic method of 2-methyl pyrroline | |
| CN115197237B (en) | Method for purifying fluorescein isomer | |
| CN115894521B (en) | Fluorescent molecule based on caged oxacalixarene, and preparation method and application thereof | |
| CN107827890A (en) | Pass through the method for the Dynamic Kinetic Resolution synthesis of chiral purine acyclonucleosides of purine, aldehyde and acid anhydrides | |
| CN114573659B (en) | Solid phase synthesis method of high flux protein group quantitative reagent | |
| CN117777151B (en) | Preparation method of AF594TSA | |
| CN114671842B (en) | Fluorescent compound, preparation method thereof, fluorescent modified nucleotide and kit | |
| CN114524770B (en) | Bis-benzimidazole fluorescent dye, preparation method and application thereof | |
| CN110028469B (en) | Preparation method and application of key intermediate of non-opioid analgesic | |
| CN109897049B (en) | Copper ion probe fused with carbazole and rhodamine structures and preparation method and application thereof | |
| WO2020102980A1 (en) | Preparation method for nucleotide for sequencing | |
| CN115246866A (en) | Synthetic method of Janus green B solid phase carrier |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |