{"id":676365,"date":"2024-08-20T06:33:24","date_gmt":"2024-08-20T05:33:24","guid":{"rendered":"https:\/\/www.thermal-engineering.org\/?p=676365"},"modified":"2024-08-20T06:33:24","modified_gmt":"2024-08-20T05:33:24","slug":"cach-may-chu-ky-nhiet-thuc-hien-pcr-nhu-the-nao","status":"publish","type":"post","link":"https:\/\/www.thermal-engineering.org\/vi\/cach-may-chu-ky-nhiet-thuc-hien-pcr-nhu-the-nao\/","title":{"rendered":"C\u00e1ch m\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o"},"content":{"rendered":"

M\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o: Gi\u1ea3i th\u00edch quy tr\u00ecnh nhi\u1ec7t gi\u00fap nh\u00e2n b\u1ea3n DNA trong ph\u00f2ng th\u00ed nghi\u1ec7m, \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng r\u1ed9ng r\u00e3i trong y h\u1ecdc v\u00e0 nghi\u00ean c\u1ee9u.<\/p>\n

\"C\u00e1ch<\/p>\n

C\u00e1ch m\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o<\/h2>\n

M\u00e1y chu k\u1ef3 nhi\u1ec7t (Thermal Cycler) l\u00e0 thi\u1ebft b\u1ecb quan tr\u1ecdng trong ph\u1ea3n \u1ee9ng chu\u1ed7i polymerase (PCR – Polymerase Chain Reaction), m\u1ed9t k\u1ef9 thu\u1eadt sinh h\u1ecdc ph\u00e2n t\u1eed \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 nh\u00e2n b\u1ea3n m\u1ed9t \u0111o\u1ea1n ADN c\u1ee5 th\u1ec3. Qu\u00e1 tr\u00ecnh PCR tr\u1ea3i qua ba b\u01b0\u1edbc ch\u00ednh l\u1eb7p \u0111i l\u1eb7p l\u1ea1i nhi\u1ec1u l\u1ea7n: bi\u1ebfn t\u00ednh (denaturation), g\u1eafn m\u1ed3i (annealing), v\u00e0 k\u00e9o d\u00e0i (extension).<\/p>\n

B\u01b0\u1edbc 1: Bi\u1ebfn t\u00ednh<\/h3>\n

Trong b\u01b0\u1edbc bi\u1ebfn t\u00ednh, m\u1eabu ADN \u0111\u01b0\u1ee3c gia nhi\u1ec7t \u0111\u1ebfn nhi\u1ec7t \u0111\u1ed9 kho\u1ea3ng 95\u00b0C \u0111\u1ec3 t\u00e1ch c\u00e1c s\u1ee3i \u0111\u00f4i ADN th\u00e0nh s\u1ee3i \u0111\u01a1n. Nhi\u1ec7t \u0111\u1ed9 cao n\u00e0y l\u00e0m \u0111\u1ee9t c\u00e1c li\u00ean k\u1ebft hydro gi\u1eefa c\u00e1c baz\u01a1 nit\u01a1 c\u1ee7a c\u00e1c s\u1ee3i.<\/p>\n

B\u01b0\u1edbc 2: G\u1eafn m\u1ed3i<\/h3>\n

\u1ede b\u01b0\u1edbc n\u00e0y, nhi\u1ec7t \u0111\u1ed9 \u0111\u01b0\u1ee3c h\u1ea1 xu\u1ed1ng kho\u1ea3ng 50-65\u00b0C, cho ph\u00e9p c\u00e1c m\u1ed3i (primers) \u0111\u1eb7c hi\u1ec7u b\u00e1m v\u00e0o c\u00e1c s\u1ee3i \u0111\u01a1n ADN m\u1ee5c ti\u00eau. M\u1ed3i l\u00e0 c\u00e1c \u0111o\u1ea1n ng\u1eafn c\u1ee7a ADN ho\u1eb7c ARN, c\u00f3 nhi\u1ec7m v\u1ee5 \u0111\u00e1nh d\u1ea5u \u0111i\u1ec3m b\u1eaft \u0111\u1ea7u cho t\u1ed5ng h\u1ee3p ADN m\u1edbi.<\/p>\n

B\u01b0\u1edbc 3: K\u00e9o d\u00e0i<\/h3>\n

Nhi\u1ec7t \u0111\u1ed9 sau \u0111\u00f3 \u0111\u01b0\u1ee3c n\u00e2ng l\u00ean kho\u1ea3ng 72\u00b0C, nhi\u1ec7t \u0111\u1ed9 t\u1ed1i \u01b0u cho ho\u1ea1t \u0111\u1ed9ng c\u1ee7a enzyme DNA polymerase. Enzyme n\u00e0y s\u1ebd th\u00eam c\u00e1c nucleotide t\u1ef1 do (dATP, dTTP, dCTP, dGTP) v\u00e0o \u0111\u1ea7u 3′ c\u1ee7a m\u1ed7i \u0111o\u1ea1n m\u1ed3i, sao ch\u00e9p chu\u1ed7i ADN m\u1edbi t\u1eeb m\u1ea1ch khu\u00f4n.<\/p>\n

Chu k\u1ef3 nhi\u1ec7t<\/h3>\n

C\u00e1c b\u01b0\u1edbc n\u00e0y \u0111\u01b0\u1ee3c l\u1eb7p l\u1ea1i t\u1eeb 20 \u0111\u1ebfn 40 chu k\u1ef3, t\u1ea1o ra h\u00e0ng tri\u1ec7u \u0111\u1ebfn h\u00e0ng t\u1ef7 b\u1ea3n sao c\u1ee7a \u0111o\u1ea1n ADN m\u1ee5c ti\u00eau t\u1eeb m\u1ed9t l\u01b0\u1ee3ng ban \u0111\u1ea7u r\u1ea5t nh\u1ecf.<\/p>\n

S\u01a1 \u0111\u1ed3 t\u00f3m t\u1eaft<\/h3>\n
    \n
  1. Bi\u1ebfn t\u00ednh: 95\u00b0C<\/li>\n
  2. G\u1eafn m\u1ed3i: 50-65\u00b0C<\/li>\n
  3. K\u00e9o d\u00e0i: 72\u00b0C<\/li>\n<\/ol>\n

    \u1ee8ng d\u1ee5ng c\u1ee7a PCR<\/h3>\n
      \n
    • Ch\u1ea9n \u0111o\u00e1n b\u1ec7nh: Ph\u00e1t hi\u1ec7n virus ho\u1eb7c vi khu\u1ea9n trong m\u1eabu b\u1ec7nh ph\u1ea9m.<\/li>\n
    • Ph\u00e2n t\u00edch ADN: Nh\u00e2n b\u1ea3n ADN trong nghi\u00ean c\u1ee9u gen.<\/li>\n
    • K\u1ef9 thu\u1eadt ph\u00e1p y: Ph\u00e2n t\u00edch b\u1eb1ng ch\u1ee9ng di truy\u1ec1n t\u1eeb hi\u1ec7n tr\u01b0\u1eddng v\u1ee5 \u00e1n.<\/li>\n<\/ul>\n

      K\u1ef9 thu\u1eadt PCR \u0111\u00e3 c\u00e1ch m\u1ea1ng h\u00f3a ng\u00e0nh sinh h\u1ecdc v\u00e0 y t\u1ebf nh\u1edd kh\u1ea3 n\u0103ng nh\u00e2n b\u1ea3n nhanh ch\u00f3ng v\u00e0 ch\u00ednh x\u00e1c c\u00e1c \u0111o\u1ea1n ADN mong mu\u1ed1n.<\/p>\n","protected":false},"excerpt":{"rendered":"

      M\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o: Gi\u1ea3i th\u00edch quy tr\u00ecnh nhi\u1ec7t gi\u00fap nh\u00e2n b\u1ea3n DNA trong ph\u00f2ng th\u00ed nghi\u1ec7m, \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng r\u1ed9ng r\u00e3i trong y h\u1ecdc v\u00e0 nghi\u00ean c\u1ee9u.<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[132],"tags":[],"yoast_head":"\nC\u00e1ch m\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o<\/title>\n<meta name=\"description\" content=\"M\u00e1y chu k\u1ef3 nhi\u1ec7t th\u1ef1c hi\u1ec7n PCR nh\u01b0 th\u1ebf n\u00e0o: Gi\u1ea3i th\u00edch quy tr\u00ecnh nhi\u1ec7t gi\u00fap nh\u00e2n b\u1ea3n DNA trong ph\u00f2ng th\u00ed nghi\u1ec7m, \u0111\u01b0\u1ee3c \u1ee9ng d\u1ee5ng r\u1ed9ng r\u00e3i trong y h\u1ecdc v\u00e0 nghi\u00ean c\u1ee9u.\" \/>\n<meta name=\"robots\" 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