{"id":2911,"date":"2023-11-14T06:09:08","date_gmt":"2023-11-14T06:09:08","guid":{"rendered":"https:\/\/oligox.abtvn.com\/?p=2911"},"modified":"2023-12-01T03:27:12","modified_gmt":"2023-12-01T03:27:12","slug":"oligonucleotide-bien-doi-thiol","status":"publish","type":"post","link":"https:\/\/oligox.abtvn.com\/en\/oligonucleotide-bien-doi-thiol\/","title":{"rendered":"OLIGONUCLEOTIDE BI\u1ebeN \u0110\u1ed4I THIOL<\/strong>"},"content":{"rendered":"

S\u1ef1 li\u00ean k\u1ebft c\u1ee7a c\u00e1c enzyme, \u0111\u1eb7c bi\u1ec7t gi\u1eefa alkaline phosphatase v\u00e0 horseradish peroxidase v\u1edbi oligo \u0111\u00f3ng vai tr\u00f2 quan tr\u1ecdng trong qu\u00e1 tr\u00ecnh nghi\u00ean c\u1ee9u, s\u1ea3n xu\u1ea5t trong ch\u1ea9n \u0111o\u00e1n probe. \u0110i\u1ec1u n\u00e0y c\u00f9ng v\u1edbi s\u1ef1 quan t\u00e2m trong \u0111\u00e1nh d\u1ea5u c\u00e1c nh\u00e3n \u0111\u1eb7c th\u00f9 cho thiol (thiol-specific tag) g\u00f3p ph\u1ea7n \u0111\u01b0a ra c\u00e1c \u00fd t\u01b0\u1edfng nghi\u00ean c\u1ee9u v\u1ec1 ph\u01b0\u01a1ng ph\u00e1p s\u1ea3n xu\u1ea5t oligonucleotide bi\u1ebfn \u0111\u1ed5i thiol (Thiol-modified oligonucleotide) trong nghi\u00ean c\u1ee9u.<\/p>\n\n\n\n

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Nh\u00f3m Thiol bi\u1ebfn \u0111\u1ed5i (Thiol-modifier) trong s\u1ea3n xu\u1ea5t c\u00e1c nh\u00f3m sulfhydryl \u1edf v\u1ecb tr\u00ed 5′-terminus \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng d\u1ef1a tr\u00ean vi\u1ec7c kh\u00f3a nh\u00f3m thiol b\u1eb1ng nh\u00f3m b\u1ea3o v\u1ec7 trityl trong qu\u00e1 tr\u00ecnh t\u1ed5ng h\u1ee3p. M\u1eb7c d\u00f9 quy tr\u00ecnh n\u00e0y \u0111\u00e3 \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng th\u00e0nh c\u00f4ng nh\u01b0ng v\u1eabn t\u1ed3n t\u1ea1i m\u1ed9t s\u1ed1 v\u1ea5n \u0111\u1ec1, nh\u01b0 m\u1ee9c \u0111\u1ed9 kh\u1eed DMT (Oxidative detritylation) th\u1ea5p trong qu\u00e1 tr\u00ecnh t\u1ed5ng h\u1ee3p v\u00e0 s\u1eed d\u1ee5ng b\u1ea1c nitrate trong b\u01b0\u1edbc kh\u1eed b\u1ea3o v\u1ec7 cu\u1ed1i c\u00f9ng.<\/p>\n\n\n\n

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B\u00e0i nghi\u00ean c\u1ee9u c\u1ee7a A. Kumar \u0111\u1ec1 c\u1eadp \u0111\u1ebfn vi\u1ec7c d\u00f9ng N-acetyl-DL-homocystein thiolactone \u0111\u1ec3 \u0111i\u1ec1u ch\u1ec9nh m\u1ed9t 5′-amino-oligonucleotide th\u00e0nh m\u1ed9t thiol. Thiol-modified oligonucleotide t\u1ea1i v\u1ecb tr\u00ed 3′-thiols \u0111\u01b0\u1ee3c t\u1ed5ng h\u1ee3p d\u1ef1a tr\u00ean c\u00e1c gi\u00e1 \u0111\u1ee1 th\u1ec3 r\u1eafn (solid support) ch\u1ee9a li\u00ean k\u1ebft disulfide. Nh\u1eefng oligonucleotide n\u00e0y bi\u1ebfn \u0111\u1ed5i thiol \u1edf v\u1ecb tr\u00ed 3′ \u0111\u1eb7c bi\u1ec7t h\u1eefu \u00edch khi mu\u1ed1n s\u1eed d\u1ee5ng m\u1ed9t lo\u1ea1i \u0111\u00e1nh d\u1ea5u kh\u00e1c cho v\u1ecb tr\u00ed \u0111\u1ea7u 5′ c\u1ee7a ch\u00fang.<\/p>\n\n\n\n

\"H\u00ecnh
H\u00ecnh 1. C\u1ea5u tr\u00fac Thiol-modifier S-S<\/strong>
(Ngu\u1ed3n: Glen Research)<\/em><\/figcaption><\/figure>\n\n\n\n
<\/div>\n\n\n\n

1. Nh\u00f3m Thiol t\u1ea1i V\u1ecb tr\u00ed 5′-Terminus<\/strong><\/h2>\n\n\n\n

a) T\u1ed5ng h\u1ee3p DMT-OFF<\/strong><\/h3>\n\n\n\n

Thiol-Modifier C6 S-S \u0111\u01b0\u1ee3c th\u00eam v\u00e0o v\u1ecb tr\u00ed \u0111\u1ea7u 5′ c\u1ee7a oligonucleotide b\u1eb1ng ph\u01b0\u01a1ng ph\u00e1p t\u1ed5ng h\u1ee3p t\u1ef1 \u0111\u1ed9ng DMT-OFF. Vi\u1ec7c gi\u1ea3i ph\u00f3ng DMT t\u1eeb chu k\u1ef3 cu\u1ed1i c\u00f3 th\u1ec3 d\u00f9ng \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh hi\u1ec7u su\u1ea5t gh\u00e9p n\u1ed1i. \u0110\u1ed1i v\u1edbi ammonium hydroxide ti\u00eau chu\u1ea9n \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 kh\u1eed b\u1ea3o v\u1ec7, dithiothreitol (DTT) \u0111\u01b0\u1ee3c th\u00eam v\u00e0o \u0111\u1ec3 \u0111\u1ea1t n\u1ed3ng \u0111\u1ed9 0,05M. Ti\u1ebfn h\u00e0nh qu\u00e1 tr\u00ecnh kh\u1eed b\u1ea3o v\u1ec7 (55\u00b0C\/16h). Quy tr\u00ecnh n\u00e0y s\u1ebd gi\u00fap lo\u1ea1i b\u1ecf c\u00e1c nh\u00f3m b\u1ea3o v\u1ec7 base v\u00e0 l\u00e0m \u0111\u1ee9t g\u00e3y li\u00ean k\u1ebft disulfide \u0111\u1ec3 t\u1ea1o ra g\u1ed1c 5′-thiol. Sau \u0111\u00f3, c\u00e1c b\u01b0\u1edbc ly t\u00e2m, lo\u1ea1i b\u1ecf mu\u1ed1i, v\u00e0 tinh s\u1ea1ch oligonucleotide bi\u1ebfn \u0111\u1ed5i thiol \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n d\u1ef1a tr\u00ean c\u00e1c quy tr\u00ecnh ti\u00eau chu\u1ea9n (n\u1ebfu c\u1ea7n).<\/p>\n\n\n\n

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b) T\u1ed5ng h\u1ee3p DMT-ON<\/strong><\/h3>\n\n\n\n

Thiol-Modifier C6 S-S \u0111\u01b0\u1ee3c th\u00eam v\u00e0o v\u1ecb tr\u00ed 5′-terminus c\u1ee7a oligonucleotide b\u1eb1ng ph\u01b0\u01a1ng ph\u00e1p t\u1ed5ng h\u1ee3p t\u1ef1 \u0111\u1ed9ng DMT-ON. Ti\u1ebfn h\u00e0nh qu\u00e1 tr\u00ecnh kh\u1eed b\u1ea3o v\u1ec7 theo c\u00e1ch th\u00f4ng th\u01b0\u1eddng. Qu\u00e1 tr\u00ecnh tinh s\u1ea1ch trityl trong oligonucleotide d\u00f9ng ph\u01b0\u01a1ng ph\u00e1p tinh s\u1ea1ch b\u1eb1ng HPLC ho\u1eb7c tr\u00ean cartridge. Sau \u0111\u00f3, ti\u1ebfn h\u00e0nh bay h\u01a1i dung d\u1ecbch s\u1ea3n ph\u1ea9m cho \u0111\u1ebfn khi kh\u00f4. S\u1eed d\u1ee5ng DTT (0,04M trong dung d\u1ecbch phosphate 0,17M, pH 8) trong 16 gi\u1edd \u0111\u1ec3 c\u1eaft li\u00ean k\u1ebft disulfide. Sau \u0111\u00f3, kh\u1eed mu\u1ed1i trong oligonucleotide b\u1eb1ng ph\u01b0\u01a1ng ph\u00e1p cartridge ho\u1eb7c c\u1ed9t. (Thiol ch\u1ee9a DMT s\u1ebd v\u1eabn g\u1eafn v\u1edbi matrix lo\u1ea1i b\u1ecf mu\u1ed1i.)<\/p>\n\n\n\n

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2. Nh\u00f3m Thiol t\u1ea1i v\u1ecb tr\u00ed 3′-Terminus trong oligonucleotide<\/strong><\/h2>\n\n\n\n

Thiol-Modifier C6 S-S \u0111\u01b0\u1ee3c th\u00eam v\u00e0o b\u1ea5t k\u1ef3 gi\u00e1 \u0111\u1ee1 nucleoside n\u00e0o ho\u1eb7c s\u1eed d\u1ee5ng Thiol-Modifier C3 S-S CPG \u0111\u1ec3 t\u1ed5ng h\u1ee3p oligonucleotide mong mu\u1ed1n. \u0110\u1ed1i v\u1edbi ammonium hydroxide ti\u00eau chu\u1ea9n \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 gi\u1ea3i b\u1ea3o v\u1ec7, dithiothreitol (DTT) \u0111\u01b0\u1ee3c th\u00eam v\u00e0o \u0111\u1ec3 \u0111\u1ea1t \u0111\u01b0\u1ee3c n\u1ed3ng \u0111\u1ed9 0,05M. Sau \u0111\u00f3, ti\u1ebfn h\u00e0nh qu\u00e1 tr\u00ecnh kh\u1eed b\u1ea3o v\u1ec7 \u1edf 55\u00b0C trong 16 gi\u1edd. Quy tr\u00ecnh n\u00e0y gi\u00fap lo\u1ea1i b\u1ecf c\u00e1c nh\u00f3m b\u1ea3o v\u1ec7 base v\u00e0 l\u00e0m \u0111\u1ee9t g\u00e3y li\u00ean k\u1ebft disulfide \u0111\u1ec3 t\u1ea1o ra g\u1ed1c 3′-thiol. Sau \u0111\u00f3, c\u00e1c b\u01b0\u1edbc ly t\u00e2m, lo\u1ea1i b\u1ecf mu\u1ed1i, v\u00e0 tinh s\u1ea1ch thiol-modified oligonucleotide \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n d\u1ef1a tr\u00ean c\u00e1c quy tr\u00ecnh ti\u00eau chu\u1ea9n (n\u1ebfu c\u1ea7n).<\/p>\n\n\n\n

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L\u01b0u \u00fd: <\/strong>Thiol-modified oligonucleotide n\u00ean \u0111\u01b0\u1ee3c b\u1ea3o qu\u1ea3n trong m\u00f4i tr\u01b0\u1eddng b\u1ea5t ho\u1ea1t ho\u1eb7c trong dung d\u1ecbch ch\u1ee9a DTT (0,01M) \u0111\u1ec3 tr\u00e1nh t\u1ea1o th\u00e0nh li\u00ean k\u1ebft disulfide do qu\u00e1 tr\u00ecnh oxi h\u00f3a. N\u1ebfu c\u1ea7n thi\u1ebft, DTT c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c tr\u00edch xu\u1ea5t t\u1eeb dung d\u1ecbch b\u1eb1ng ethyl acetate. Ti\u1ebfp t\u1ee5c \u0111\u1ebfn ph\u1ea3n \u1ee9ng li\u00ean k\u1ebft.<\/p>\n\n\n\n

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Pannel \u1ee9ng d\u1ee5ng trong ch\u1ea9n \u0111o\u00e1n Thalassemia<\/a><\/h3>
\n \n \n<\/svg>\nadmin<\/a><\/span>\n \n<\/svg>\nAug 26, 2025<\/span>\n \n \n<\/svg>\n18 min read<\/span><\/div>

T\u1ed5ng quan Thalassemia Gi\u1edbi thi\u1ec7u Thalassemia Thalassemia (hay c\u00f2n \u0111\u01b0\u1ee3c g\u1ecdi v\u1edbi t\u00ean b\u1ec7nh tan m\u00e1u b\u1ea9m sinh) l\u00e0 m\u1ed9t b\u1ec7nh l\u00fd huy\u1ebft h\u1ecdc di truy\u1ec1n c\u00f3 t\u1ef7 l\u1ec7 l\u01b0u h\u00e0nh cao tr\u00ean to\u00e0n c\u1ea7u v\u00e0 l\u00e0 m\u1ed9t v\u1ea5n \u0111\u1ec1…<\/p>\n<\/div>

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\u1ea2nh h\u01b0\u1edfng c\u1ee7a TaqMan probe \u0111\u1ebfn \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u00e0 \u0111\u1ed9 nh\u1ea1y trong Realtime PCR<\/a><\/h3>
\n \n \n<\/svg>\nadmin<\/a><\/span>\n \n<\/svg>\nMay 27, 2025<\/span>\n \n \n<\/svg>\n26 min read<\/span><\/div>

TaqMan Probe \u0111\u00f3ng vai tr\u00f2 trung t\u00e2m trong vi\u1ec7c \u0111\u1ea3m b\u1ea3o \u0111\u1ed9 ch\u00ednh x\u00e1c, \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u00e0 \u0111\u1ed9 nh\u1ea1y c\u1ee7a ph\u1ea3n \u1ee9ng trong k\u1ef9 thu\u1eadt Real-time PCR<\/p>\n<\/div>

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\"TAQMAN<\/a><\/div><\/div>
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TAQMAN PROBE V\u00c0 C\u00c1C TI\u00caU CH\u00cd QUAN TR\u1eccNG KHI THI\u1ebeT K\u1ebe PROBE TRONG REAL TIME PCR<\/a><\/h3>
\n \n \n<\/svg>\nadmin<\/a><\/span>\n \n<\/svg>\nMay 20, 2025<\/span>\n \n \n<\/svg>\n12 min read<\/span><\/div>

Probe (\u0111\u1ea7u d\u00f2) trong sinh h\u1ecdc ph\u00e2n t\u1eed l\u00e0 m\u1ed9t \u0111o\u1ea1n oligonucleotide (DNA ho\u1eb7c RNA s\u1ee3i \u0111\u01a1n) ng\u1eafn, \u0111\u01b0\u1ee3c \u0111\u00e1nh d\u1ea5u b\u1eb1ng c\u00e1c ph\u00e2n t\u1eed ph\u00e1t t\u00edn hi\u1ec7u (th\u01b0\u1eddng l\u00e0 c\u00e1c ch\u1ea5t ph\u00e1t hu\u1ef3nh quang). Probe \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf \u0111\u1ec3…<\/p>\n<\/div>

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HPLC L\u00e0 G\u00ec? \u1ee8ng D\u1ee5ng C\u1ee7a HPLC Trong Tinh S\u1ea1ch v\u00e0 Ph\u00e2n T\u00edch Oligonucleotide<\/a><\/h3>
\n \n \n<\/svg>\nadmin<\/a><\/span>\n \n<\/svg>\nFeb 14, 2025<\/span>\n \n \n<\/svg>\n13 min read<\/span><\/div>

HPLC (High Performance Liquid Chromatography) \u2013 S\u1eafc k\u00fd l\u1ecfng hi\u1ec7u n\u0103ng cao l\u00e0 m\u1ed9t k\u1ef9 thu\u1eadt ph\u00e2n t\u00edch \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 t\u00e1ch, \u0111\u1ecbnh t\u00ednh ho\u1eb7c \u0111\u1ecbnh l\u01b0\u1ee3ng c\u00e1c ch\u1ea5t, th\u00e0nh ph\u1ea7n trong m\u1ed9t h\u1ed7n h\u1ee3p ch\u1ea5t l\u1ecfng.<\/p>\n<\/div>

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C\u00c1C PH\u01af\u01a0NG PH\u00c1P TINH S\u1ea0CH OLIGONUCLEOTIDE<\/a><\/h3>
\n \n \n<\/svg>\nadmin<\/a><\/span>\n \n<\/svg>\nJan 15, 2025<\/span>\n \n \n<\/svg>\n9 min read<\/span><\/div>

Oligonucleotide l\u00e0 nh\u1eefng ph\u00e2n t\u1eed DNA ho\u1eb7c RNA m\u1ea1ch \u0111\u01a1n ho\u1eb7c m\u1ea1ch \u0111\u00f4i c\u00f3 k\u00edch th\u01b0\u1edbc ng\u1eafn th\u01b0\u1eddng \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng trong c\u00e1c \u1ee9ng d\u1ee5ng sinh h\u1ecdc ph\u00e2n t\u1eed, y h\u1ecdc v\u00e0 nghi\u00ean c\u1ee9u khoa h\u1ecdc. C\u00e1c tr\u00ecnh t\u1ef1 oligonucleotide…<\/p>\n<\/div>

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D\u1ecbch v\u1ee5 OligoX<\/a><\/em><\/strong><\/h3>

T\u1ed5ng h\u1ee3p Oligonucleotide bao g\u1ed3m c\u00e1c s\u1ea3n ph\u1ea9m PCR, qPCR Primer, Primer\/Probe bi\u1ebfn \u0111\u1ed5i, Tr\u00ecnh t\u1ef1 RNA, Amplicon v\u00e0 c\u00e1c t\u1ed5ng h\u1ee3p \u0111\u1eb7c bi\u1ec7t<\/p><\/div>

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Ngu\u1ed3n tham kh\u1ea3o<\/strong><\/h2>\n\n\n\n
    \n
  1. Glen Research. https:\/\/www.glenresearch.com\/reports\/gr5-15<\/a><\/li>\n\n\n\n
  2. B.A. Connolly and R. Rider, Nucleic Acids Res., 1985, 13, 4485.<\/li>\n\n\n\n
  3. B.A. Connolly, Nucleic Acids Res., 1987, 15, 3131-3139.<\/li>\n\n\n\n
  4. N.D. Sinha and R.M. Cook, Nucleic Acids Res., 1988, 16, 2659.<\/li>\n\n\n\n
  5. A. Kumar, S. Advani, H. Dawar, and G.P. Talwar, Nucleic Acids Res., 1991, 19, 4561.<\/li>\n\n\n\n
  6. R. Zuckermann, D. Corey, and P. Schultz, Nucleic Acids Res., 1987, 15, 5305.<\/li>\n\n\n\n
  7. K.C. Gupta, P. Shanna, S. Sathyanarayana, and P. Kumar, Tetrahedron Lett., 1990, 31, 2471-2474.<\/li>\n\n\n\n
  8. U. Asseline, E. Bonfils, R. Kurfurst, M. Chassignol, V. Roig, and N.T. Thuong, Tetrahedron, 1992, 48, 1233-1254.<\/li>\n<\/ol>","protected":false},"excerpt":{"rendered":"

    S\u1ef1 li\u00ean k\u1ebft c\u1ee7a c\u00e1c enzyme, \u0111\u1eb7c bi\u1ec7t gi\u1eefa alkaline phosphatase v\u00e0 horseradish peroxidase v\u1edbi oligo \u0111\u00f3ng vai tr\u00f2 quan tr\u1ecdng trong qu\u00e1 tr\u00ecnh nghi\u00ean c\u1ee9u, s\u1ea3n xu\u1ea5t trong ch\u1ea9n \u0111o\u00e1n probe. \u0110i\u1ec1u n\u00e0y c\u00f9ng v\u1edbi s\u1ef1 quan t\u00e2m trong \u0111\u00e1nh d\u1ea5u c\u00e1c nh\u00e3n \u0111\u1eb7c th\u00f9 cho thiol (thiol-specific tag) g\u00f3p ph\u1ea7n \u0111\u01b0a ra c\u00e1c …<\/p>\n

    OLIGONUCLEOTIDE BI\u1ebeN \u0110\u1ed4I THIOL<\/strong><\/span> Read More 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li\u00ean k\u1ebft c\u1ee7a c\u00e1c enzyme, \u0111\u1eb7c bi\u1ec7t gi\u1eefa alkaline phosphatase v\u00e0 horseradish peroxidase v\u1edbi oligo \u0111\u00f3ng vai tr\u00f2 quan tr\u1ecdng trong qu\u00e1 tr\u00ecnh nghi\u00ean c\u1ee9u, s\u1ea3n xu\u1ea5t trong ch\u1ea9n \u0111o\u00e1n probe. \u0110i\u1ec1u n\u00e0y c\u00f9ng v\u1edbi s\u1ef1 quan t\u00e2m trong \u0111\u00e1nh d\u1ea5u c\u00e1c nh\u00e3n \u0111\u1eb7c th\u00f9 cho thiol (thiol-specific tag) g\u00f3p ph\u1ea7n \u0111\u01b0a ra 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