{"id":11743,"date":"2025-01-15T12:16:28","date_gmt":"2025-01-15T04:46:28","guid":{"rendered":"https:\/\/abtvn.com\/?p=11743"},"modified":"2025-01-22T10:12:26","modified_gmt":"2025-01-22T02:42:26","slug":"khuech-dai-doan-san-pham-ngan-trong-real-time-pcr","status":"publish","type":"post","link":"https:\/\/abtvn.com\/en\/khuech-dai-doan-san-pham-ngan-trong-real-time-pcr\/","title":{"rendered":"V\u00ec sao n\u00ean l\u1ef1a ch\u1ecdn khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn trong Real-time PCR?<\/strong>"},"content":{"rendered":"

Real-time PCR (qPCR)<\/strong> cho ph\u00e9p x\u00e1c \u0111\u1ecbnh l\u01b0\u1ee3ng DNA sau m\u1ed7i chu k\u1ef3 th\u00f4ng qua thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang t\u1ea1o ra t\u00edn hi\u1ec7u hu\u1ef3nh quang t\u0103ng d\u1ea7n theo t\u1ef7 l\u1ec7 thu\u1eadn v\u1edbi s\u1ed1 l\u01b0\u1ee3ng s\u1ea3n ph\u1ea9m PCR (amplicon)<\/strong> \u0111\u01b0\u1ee3c t\u1ea1o ra (Lela Buckingham, Maribeth Flaws, 2007)<\/em>. M\u1ed9t trong nh\u1eefng y\u1ebfu t\u1ed1 quan tr\u1ecdng quy\u1ebft \u0111\u1ecbnh s\u1ef1 th\u00e0nh c\u00f4ng c\u1ee7a m\u1ed9t ph\u1ea3n \u1ee9ng PCR ch\u00ednh l\u00e0 thi\u1ebft k\u1ebf \u0111o\u1ea1n khu\u1ebfch \u0111\u1ea1i (amplicon) v\u1edbi \u0111\u1ed9 d\u00e0i s\u1ea3n ph\u1ea9m ph\u00f9 h\u1ee3p.<\/p>\n\n\n\n

Vi\u1ec7c l\u1ef1a ch\u1ecdn m\u1ed9t \u0111o\u1ea1n khu\u1ebfch \u0111\u1ea1i ph\u00f9 h\u1ee3p kh\u00f4ng ch\u1ec9 \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn \u0111\u1ed9 nh\u1ea1y, \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng m\u00e0 c\u00f2n quy\u1ebft \u0111\u1ecbnh \u0111\u1ebfn ch\u1ea5t l\u01b0\u1ee3ng v\u00e0 \u0111\u1ed9 tin c\u1eady c\u1ee7a k\u1ebft qu\u1ea3 thu \u0111\u01b0\u1ee3c. V\u1eady v\u00ec sao ch\u00fang ta th\u01b0\u1eddng h\u01b0\u1edbng \u0111\u1ebfn vi\u1ec7c thi\u1ebft k\u1ebf c\u00e1c \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn? B\u00e0i vi\u1ebft n\u00e0y s\u1ebd \u0111i s\u00e2u v\u00e0o l\u00fd gi\u1ea3i nh\u1eefng l\u1ee3i \u00edch c\u1ee7a vi\u1ec7c n\u00e0y, \u0111\u1ed3ng th\u1eddi tr\u00ecnh b\u00e0y c\u00e1c y\u1ebfu t\u1ed1 c\u1ea7n c\u00e2n nh\u1eafc khi thi\u1ebft k\u1ebf primer.<\/p>\n\n\n\n

1. T\u1ed5ng quan v\u1ec1 Real-time PCR<\/strong><\/h2>\n\n\n\n

PCR (Polymerase Chain Reaction)<\/strong> l\u00e0 m\u1ed9t c\u00f4ng c\u1ee5 m\u1ea1nh m\u1ebd trong nghi\u00ean c\u1ee9u sinh h\u1ecdc, \u0111\u01b0\u1ee3c s\u1eed d\u1ee5ng \u0111\u1ec3 khu\u1ebfch \u0111\u1ea1i m\u1ed9t \u0111o\u1ea1n DNA \u0111\u1eb7c hi\u1ec7u t\u1eeb m\u1ed9t m\u1eabu ban \u0111\u1ea7u c\u00f3 ch\u1ee9a l\u01b0\u1ee3ng DNA r\u1ea5t nh\u1ecf do Karl Mullis<\/em> v\u00e0 c\u00e1c c\u00f4ng s\u1ef1 nghi\u00ean c\u1ee9u ph\u00e1t tri\u1ec3n v\u00e0o n\u0103m 1985 (Huong, 2003)<\/em>. PCR ho\u1ea1t \u0111\u1ed9ng d\u1ef1a tr\u00ean vi\u1ec7c m\u00f4 ph\u1ecfng qu\u00e1 tr\u00ecnh nh\u00e2n \u0111\u00f4i DNA \u1edf t\u1ebf b\u00e0o, theo \u0111\u00f3 vi\u1ec7c l\u1eb7p l\u1ea1i c\u00e1c chu k\u1ef3 nhi\u1ec7t bao g\u1ed3m c\u00e1c b\u01b0\u1edbc bi\u1ebfn t\u00ednh, b\u1eaft c\u1eb7p primer v\u00e0 k\u00e9o d\u00e0i chu\u1ed7i nucleic acid cho ph\u00e9p t\u1ea1o ra s\u1ed1 l\u01b0\u1ee3ng l\u1edbn c\u00e1c b\u1ea3n sao c\u1ee7a \u0111o\u1ea1n DNA \u0111\u00edch.<\/p>\n\n\n\n

\u0110\u1ebfn nay, PCR \u0111ang d\u1ea7n \u0111\u01b0\u1ee3c xem l\u00e0 \u201cti\u00eau chu\u1ea9n v\u00e0ng\u201d trong nhi\u1ec1u x\u00e9t nghi\u1ec7m ch\u1ea9n \u0111o\u00e1n b\u1ec7nh nh\u1edd v\u00e0o \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u cao. Tuy nhi\u00ean, PCR truy\u1ec1n th\u1ed1ng c\u00f2n nhi\u1ec1u h\u1ea1n ch\u1ebf nh\u01b0 ph\u1ea3i th\u1ef1c \u0111i\u1ec7n di h\u1eadu PCR \u0111\u1ec3 ph\u00e2n t\u00edch k\u1ebft qu\u1ea3, ch\u1ec9 c\u00f3 th\u1ec3 \u0111\u1ecbnh t\u00ednh s\u1ef1 hi\u1ec7n di\u1ec7n c\u1ee7a t\u00e1c nh\u00e2n, \u0111\u1ed3ng th\u1eddi ti\u1ec1m \u1ea9n nguy c\u01a1 ngo\u1ea1i nhi\u1ec5m cao, \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn c\u00e1c ph\u1ea3n \u1ee9ng ti\u1ebfn h\u00e0nh sau \u0111\u00f3.<\/p>\n\n\n\n

Ngo\u00e0i \u0111\u1ecbnh t\u00ednh, nhi\u1ec1u x\u00e9t nghi\u1ec7m hi\u1ec7n nay \u0111\u00f2i h\u1ecfi \u0111\u00f2i h\u1ecfi k\u1ebft qu\u1ea3 ph\u00e2n t\u00edch \u0111\u1ecbnh l\u01b0\u1ee3ng. Do \u0111\u00f3, Real-time PCR (PCR th\u1eddi gian th\u1ef1c) \u0111\u01b0\u1ee3c ra \u0111\u1eddi, v\u1eeba ph\u00e1t huy \u0111\u01b0\u1ee3c \u01b0u \u0111i\u1ec3m c\u1ee7a k\u1ef9 thu\u1eadt PCR truy\u1ec1n th\u1ed1ng v\u1eeba kh\u1eafc ph\u1ee5c \u0111\u01b0\u1ee3c c\u00e1c h\u1ea1n ch\u1ebf t\u1ed3n \u0111\u1ecdng mang l\u1ea1i nhi\u1ec1u \u01b0u \u0111i\u1ec3m v\u01b0\u1ee3t tr\u1ed9i.<\/p>\n\n\n\n

\"H\u00ecnh
Figure 1. <\/strong>Quy tr\u00ecnh Real-time PCR (ABT, 2023)<\/em><\/figcaption><\/figure>\n\n\n\n

2. Nh\u1eefng l\u00fd do v\u00ec sao n\u00ean khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn trong Real-time PCR?<\/strong><\/h2>\n\n\n\n

So v\u1edbi s\u1ea3n ph\u1ea9m PCR truy\u1ec1n th\u1ed1ng c\u00f3 k\u00edch th\u01b0\u1edbc t\u1eeb 200 \u2013 1000 bp, ti\u00eau chu\u1ea9n amplicon c\u1ee7a Real-time PCR \u0111\u01b0\u1ee3c thi\u1ebft l\u1eadp trong kho\u1ea3ng t\u1eeb 80 \u2013 150 bp (IDT, 2024)<\/em>. \u0110i\u1ec1u n\u00e0y d\u1eabn \u0111\u1ebfn c\u00e1c \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn trong Real-time PCR \u0111\u01b0\u1ee3c khu\u1ebfch \u0111\u1ea1i m\u1ed9t c\u00e1ch nhanh ch\u00f3ng v\u00e0 hi\u1ec7u qu\u1ea3 h\u01a1n. Ngo\u00e0i ra, s\u1ea3n ph\u1ea9m ng\u1eafn c\u00f2n h\u1ea1n ch\u1ebf \u0111\u01b0\u1ee3c xu h\u01b0\u1edbng h\u00ecnh th\u00e0nh c\u00e1c c\u1ea5u tr\u00fac th\u1ee9 c\u1ea5p nh\u01b0 k\u1eb9p t\u00f3c (hairpin) ho\u1eb7c c\u00e1c c\u1ea5u tr\u00fac g\u1ea5p kh\u00e1c, g\u00e2y c\u1ea3n tr\u1edf qu\u00e1n tr\u00ecnh nh\u00e2n b\u1ea3n. Nh\u1edd v\u1eady, qPCR \u0111\u01b0\u1ee3c t\u0103ng c\u01b0\u1eddng hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i v\u00e0 \u0111\u1ea1t hi\u1ec7u qu\u1ea3 cao h\u01a1n (Jain, 2020)<\/em>.<\/p>\n\n\n\n

Ngo\u00e0i ra, nh\u00e2n b\u1ea3n c\u00e1c \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn th\u01b0\u1eddng di\u1ec5n ra nhanh h\u01a1n so v\u1edbi c\u00e1c \u0111o\u1ea1n s\u1ea3n ph\u1ea9m d\u00e0i.\u00a0\u0110i\u1ec1u n\u00e0y gi\u00fap ti\u1ebft ki\u1ec7m th\u1eddi gian v\u00e0 t\u00e0i nguy\u00ean trong ph\u00f2ng th\u00ed nghi\u1ec7m, \u0111\u1ea3m b\u1ea3o t\u00ednh kinh t\u1ebf, \u0111\u1ed3ng th\u1eddi t\u0103ng c\u01b0\u1eddng hi\u1ec7u qu\u1ea3 c\u00f4ng vi\u1ec7c<\/a>. Trong nghi\u00ean c\u1ee9u \u0111\u1ecbnh l\u01b0\u1ee3ng, khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn gi\u00fap t\u0103ng c\u01b0\u1eddng kh\u1ea3 n\u0103ng chu\u1ea9n h\u00f3a v\u00e0 so s\u00e1nh gi\u1eefa c\u00e1c m\u1eabu kh\u00e1c nhau, t\u1eeb \u0111\u00f3, c\u00e1c k\u1ebft lu\u1eadn khoa h\u1ecdc \u0111\u01b0\u1ee3c \u0111\u01b0a ra m\u1ed9t c\u00e1ch ch\u00ednh x\u00e1c (Lela Buckingham, Maribeth Flaws, 2007)<\/em>.<\/p>\n\n\n\n

\u0110\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng qPCR lu\u00f4n \u0111\u01b0\u1ee3c ch\u00fa tr\u1ecdng v\u00e0 t\u0103ng c\u01b0\u1eddng. Do \u0111\u00f3, vi\u1ec7c khu\u1ebfch \u0111\u1ea1i c\u00e1c \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn trong qPCR gi\u00fap gi\u1ea3m thi\u1ec3u \u1ea3nh h\u01b0\u1edfng b\u1edfi c\u00e1c y\u1ebfu t\u1ed1 \u1ee9c ch\u1ebf trong m\u1eabu, nh\u01b0 c\u00e1c protein, h\u1ee3p ch\u1ea5t h\u00f3a h\u1ecdc\u2026 V\u00ec v\u1eady, \u0111\u1ed9 ch\u00ednh x\u00e1c v\u00e0 \u0111\u1ed9 nh\u1ea1y c\u1ee7a ph\u1ea3n \u1ee9ng \u0111\u01b0\u1ee3c n\u00e2ng cao, \u0111\u1eb7c bi\u1ec7t l\u00e0 khi ph\u00e2n t\u00edch c\u00e1c m\u1eabu ph\u1ee9c t\u1ea1p nh\u01b0 m\u1eabu m\u00f4i tr\u01b0\u1eddng ho\u1eb7c m\u1eabu l\u00e2m s\u00e0ng.<\/p>\n\n\n\n

3. Khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a Real-time PCR<\/strong><\/h2>\n\n\n\n

\u0110\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng qPCR li\u00ean quan m\u1eadt thi\u1ebft \u0111\u1ebfn vi\u1ec7c thi\u1ebft k\u1ebf khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn. \u0110\u1ed9 nh\u1ea1y c\u1ee7a ph\u1ea3n \u1ee9ng li\u00ean quan \u0111\u1ebfn k\u00edch th\u01b0\u1edbc c\u1ee7a c\u00e1c \u0111o\u1ea1n \u0111\u01b0\u1ee3c khu\u1ebfch \u0111\u1ea1i. C\u00e1c m\u1ee5c ti\u00eau trong Real-time PCR c\u00f3 th\u1ec3 r\u1ea5t ng\u1eafn v\u00e0 th\u01b0\u1eddng kh\u00f4ng v\u01b0\u1ee3t qu\u00e1 150 bp, nhi\u1ec1u nghi\u00ean c\u1ee9u ch\u1ec9 ra r\u1eb1ng, hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i \u0111\u1ea1t kho\u1ea3ng 100% \u0111\u1ed1i v\u1edbi c\u00e1c s\u1ea3n ph\u1ea9m k\u00edch th\u01b0\u1edbc ng\u1eafn, c\u00e1c amplicon c\u00e0ng d\u00e0i, hi\u1ec7u qu\u1ea3 c\u00f3 xu h\u01b0\u1edbng gi\u1ea3m d\u1ea7n (Debode, F., Marien, A., Janssen, \u00c9., Bragard, C., & Berben, G, 2017)<\/em><\/p>\n\n\n\n

\u0110i\u1ec1u n\u00e0y t\u1ea1o n\u00ean s\u1ef1 v\u01b0\u1ee3t tr\u1ed9i so v\u1edbi PCR truy\u1ec1n th\u1ed1ng b\u1edfi c\u00e1c \u0111o\u1ea1n th\u01b0\u1eddng d\u00e0i h\u01a1n \u0111\u1ec3 \u0111\u1ea3m b\u1ea3o d\u1ec5 h\u00ecnh dung h\u01a1n tr\u00ean gel agarose. Vi\u1ec7c gi\u1edbi h\u1ea1n \u0111\u1ed9 d\u00e0i \u0111o\u1ea1n khu\u1ebfch \u0111\u1ea1i trong kho\u1ea3ng t\u1eeb 80 \u2013 150 bp gi\u00fap r\u00fat ng\u1eafn th\u1eddi gian g\u1eafn m\u1ed3i v\u00e0 k\u00e9o d\u00e0i c\u1ee7a ph\u1ea3n \u1ee9ng v\u00e0 h\u1ea1n ch\u1ebf nh\u1eefng \u1ea3nh h\u01b0\u1edfng ti\u00eau c\u1ef1c \u0111\u1ebfn \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng qPCR ch\u1ee7 y\u1ebfu do li\u00ean k\u1ebft kh\u00f4ng chuy\u00ean bi\u1ec7t c\u1ee7a c\u00e1c primer, bao g\u1ed3m li\u00ean k\u1ebft v\u1edbi c\u00e1c tr\u00ecnh t\u1ef1 t\u01b0\u01a1ng t\u1ef1 trong DNA khu\u00f4n \u1edf c\u00e1c v\u1ecb tr\u00ed ng\u1eabu nhi\u00ean, li\u00ean k\u1ebft gi\u1eefa c\u00e1c primer kh\u00e1c nhau v\u00e0 primer t\u1ef1 li\u00ean k\u1ebft. B\u00ean c\u1ea1nh \u0111\u00f3, vi\u1ec7c thi\u1ebft k\u1ebf c\u00e1c \u0111o\u1ea1n m\u1ed3i d\u00e0i kho\u1ea3ng 18 \u2013 24 nucleotide, v\u1edbi Tm kho\u1ea3ng 65 \u00b0C gi\u00fap l\u00e0m gi\u1ea3m kh\u1ea3 n\u0103ng c\u00e1c \u0111o\u1ea1n m\u1ed3i li\u00ean k\u1ebft sai v\u1ecb tr\u00ed . \u0110\u1ed3ng th\u1eddi, s\u1eed d\u1ee5ng DNA polymerase mang c\u00f4ng ngh\u1ec7 \u201chot start\u201d s\u1ebd h\u1ea1n ch\u1ebf \u0111\u00e1ng k\u1ec3 kh\u1ea3 n\u0103ng h\u00ecnh th\u00e0nh c\u00e1c primer-dimer, v\u00ec kh\u00f4ng c\u00f3 s\u1ef1 khu\u1ebfch \u0111\u1ea1i \u201cr\u00f2 r\u1ec9\u201d n\u00e0o \u0111\u01b0\u1ee3c b\u1eaft \u0111\u1ea7u tr\u01b0\u1edbc khi c\u00e1c m\u1ed3i li\u00ean k\u1ebft v\u1edbi c\u00e1c v\u1ecb tr\u00ed tr\u00ecnh t\u1ef1 th\u00edch h\u1ee3p.<\/p>\n\n\n\n

\"\"
Figure 2 <\/strong>\u0110\u01b0\u1eddng cong khu\u1ebfch \u0111\u1ea1i thu \u0111\u01b0\u1ee3c tr\u00ean c\u00e1c amplicon c\u00f3 k\u00edch th\u01b0\u1edbc t\u0103ng d\u1ea7n (Debode, F., Marien, A., Janssen, \u00c9., Bragard, C., & Berben, G, 2017)<\/figcaption><\/figure>\n\n\n\n

4. H\u1ea1n ch\u1ebf trong thi\u1ebft k\u1ebf \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn Real-time PCR<\/strong><\/h2>\n\n\n\n

Thi\u1ebft k\u1ebf khu\u1ebfch \u0111\u1ea1i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m ng\u1eafn trong Real-time PCR mang l\u1ea1i nhi\u1ec1u l\u1ee3i \u00edch quan tr\u1ecdng, t\u1eeb vi\u1ec7c t\u0103ng c\u01b0\u1eddng hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i, gi\u1ea3m thi\u1ec3u s\u1ef1 h\u00ecnh th\u00e0nh c\u1ea5u tr\u00fac th\u1ee9 c\u1ea5p, t\u0103ng c\u01b0\u1eddng \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u, ti\u1ebft ki\u1ec7m th\u1eddi gian v\u00e0 t\u00e0i nguy\u00ean, \u0111\u1ebfn vi\u1ec7c t\u0103ng c\u01b0\u1eddng kh\u1ea3 n\u0103ng ph\u00e1t hi\u1ec7n c\u00e1c bi\u1ebfn th\u1ec3 nh\u1ecf v\u00e0 gi\u1ea3m thi\u1ec3u t\u00e1c \u0111\u1ed9ng c\u1ee7a c\u00e1c y\u1ebfu t\u1ed1 \u1ee9c ch\u1ebf. Nh\u1eefng l\u1ee3i \u00edch n\u00e0y gi\u00fap \u0111\u1ea3m b\u1ea3o r\u1eb1ng ph\u1ea3n \u1ee9ng qPCR di\u1ec5n ra m\u1ed9t c\u00e1ch ch\u00ednh x\u00e1c, hi\u1ec7u qu\u1ea3 v\u00e0 \u0111\u00e1ng tin c\u1eady, t\u1eeb \u0111\u00f3 h\u1ed7 tr\u1ee3 t\u1ed1t h\u01a1n cho c\u00e1c nghi\u00ean c\u1ee9u sinh h\u1ecdc ph\u00e2n t\u1eed v\u00e0 \u1ee9ng d\u1ee5ng y h\u1ecdc.<\/p>\n\n\n\n

M\u1eb7c d\u00f9 vi\u1ec7c thi\u1ebft k\u1ebf \u0111o\u1ea1n s\u1ea3n ph\u1ea9m PCR ng\u1eafn mang l\u1ea1i nhi\u1ec1u l\u1ee3i \u00edch, tuy nhi\u00ean n\u00f3 c\u0169ng \u0111\u1eb7t ra m\u1ed9t s\u1ed1 th\u00e1ch th\u1ee9c. M\u1ed9t trong nh\u1eefng kh\u00f3 kh\u0103n c\u00f3 th\u1ec3 k\u1ec3 \u0111\u1ebfn l\u00e0 vi\u1ec7c thi\u1ebft k\u1ebf k\u1ebf c\u1eb7p primer \u0111\u1eb7c hi\u1ec7u cho c\u00e1c v\u00f9ng l\u1eb7p l\u1ea1i trong genome. S\u1ef1 t\u01b0\u01a1ng \u0111\u1ed3ng cao gi\u1eefa c\u00e1c v\u00f9ng l\u1eb7p l\u1ea1i n\u00e0y d\u1ec5 d\u1eabn \u0111\u1ebfn hi\u1ec7n t\u01b0\u1ee3ng b\u1eaft c\u1eb7p kh\u00f4ng \u0111\u1eb7c hi\u1ec7u c\u1ee7a primer, g\u00e2y ra c\u00e1c s\u1ea3n ph\u1ea9m khu\u1ebfch \u0111\u1ea1i kh\u00f4ng mong mu\u1ed1n. B\u00ean c\u1ea1nh \u0111\u00f3, c\u00e1c bi\u1ebfn th\u1ec3 di truy\u1ec1n c\u0169ng c\u00f3 th\u1ec3 \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i, khi c\u00e1c bi\u1ebfn \u0111\u1ed5i trong tr\u00ecnh t\u1ef1 DNA l\u00e0m thay \u0111\u1ed5i v\u1ecb tr\u00ed b\u1eaft c\u1eb7p c\u1ee7a primer. Cu\u1ed1i c\u00f9ng, \u0111o\u1ea1n s\u1ea3n ph\u1ea9m PCR qu\u00e1 ng\u1eafn c\u00f3 th\u1ec3 kh\u00f4ng \u0111\u1ee7 \u1ed5n \u0111\u1ecbnh, d\u1ec5 b\u1ecb ph\u00e2n h\u1ee7y b\u1edfi c\u00e1c enzyme nuclease trong qu\u00e1 tr\u00ecnh ph\u1ea3n \u1ee9ng v\u00e0 sau ph\u1ea3n \u1ee9ng, t\u1eeb \u0111\u00f3 l\u00e0m gi\u1ea3m \u0111\u1ed9 tin c\u1eady c\u1ee7a k\u1ebft qu\u1ea3. Do \u0111\u00f3, vi\u1ec7c l\u1ef1a ch\u1ecdn \u0111\u1ed9 d\u00e0i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m PCR c\u1ea7n c\u00e2n nh\u1eafc k\u1ef9 l\u01b0\u1ee1ng \u0111\u1ec3 \u0111\u1ea3m b\u1ea3o c\u1ea3 hi\u1ec7u qu\u1ea3 khu\u1ebfch \u0111\u1ea1i v\u00e0 \u0111\u1ed9 ch\u00ednh x\u00e1c c\u1ee7a k\u1ebft qu\u1ea3.<\/p>\n\n\n\n

Kh\u00f3 kh\u0103n trong vi\u1ec7c thi\u1ebft k\u1ebf primer \u0111\u1eb7c hi\u1ec7u cho c\u00e1c v\u00f9ng l\u1eb7p l\u1ea1i trong gen 16S rRNA c\u1ee7a vi khu\u1ea9n l\u00e0 m\u1ed9t v\u00ed d\u1ee5 \u0111i\u1ec3n h\u00ecnh. B\u00ean c\u1ea1nh \u0111\u00f3, s\u1ef1 \u0111a d\u1ea1ng v\u1ec1 di truy\u1ec1n c\u1ee7a virus HIV l\u00e0m cho vi\u1ec7c thi\u1ebft k\u1ebf primer cho c\u00e1c v\u00f9ng m\u00e3 h\u00f3a protein bao ph\u1ee7 tr\u1edf n\u00ean ph\u1ee9c t\u1ea1p. \u0110\u1ec3 kh\u1eafc ph\u1ee5c nh\u1eefng h\u1ea1n ch\u1ebf n\u00e0y, c\u00e1c nh\u00e0 nghi\u00ean c\u1ee9u th\u01b0\u1eddng s\u1eed d\u1ee5ng c\u00e1c ph\u1ea7n m\u1ec1m thi\u1ebft k\u1ebf primer chuy\u00ean d\u1ee5ng nh\u01b0 Primer3, Primer-BLAST \u0111\u1ec3 \u0111\u00e1nh gi\u00e1 t\u00ednh \u0111\u1eb7c hi\u1ec7u c\u1ee7a primer v\u00e0 t\u1ed1i \u01b0u h\u00f3a \u0111i\u1ec1u ki\u1ec7n ph\u1ea3n \u1ee9ng. Ngo\u00e0i ra, vi\u1ec7c k\u1ebft h\u1ee3p PCR v\u1edbi c\u00e1c k\u1ef9 thu\u1eadt kh\u00e1c nh\u01b0 Sanger sequencing c\u00f3 th\u1ec3 gi\u00fap x\u00e1c nh\u1eadn \u0111\u1ed9 ch\u00ednh x\u00e1c c\u1ee7a k\u1ebft qu\u1ea3\u2026<\/p>\n\n\n\n

Vi\u1ec7c thi\u1ebft k\u1ebf \u0111o\u1ea1n s\u1ea3n ph\u1ea9m PCR hi\u1ec7u qu\u1ea3 \u0111\u00f2i h\u1ecfi ph\u1ea3i c\u00e2n nh\u1eafc k\u1ef9 l\u01b0\u1ee1ng nhi\u1ec1u y\u1ebfu t\u1ed1. Chi\u1ec1u d\u00e0i \u0111o\u1ea1n s\u1ea3n ph\u1ea9m th\u01b0\u1eddng dao \u0111\u1ed9ng t\u1eeb 50-150 base pair \u0111\u1ec3 \u0111\u1ea3m b\u1ea3o hi\u1ec7u su\u1ea5t khu\u1ebfch \u0111\u1ea1i t\u1ed1i \u01b0u. Nhi\u1ec7t \u0111\u1ed9 annealing ph\u00f9 h\u1ee3p l\u00e0 y\u1ebfu t\u1ed1 quy\u1ebft \u0111\u1ecbnh \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u c\u1ee7a ph\u1ea3n \u1ee9ng. Ngo\u00e0i ra, th\u00e0nh ph\u1ea7n nucleotide c\u1ee7a primer c\u1ea7n \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf c\u00e2n \u0111\u1ed1i, tr\u00e1nh c\u00e1c \u0111o\u1ea1n b\u1ed5 sung li\u00ean ti\u1ebfp \u0111\u1ec3 gi\u1ea3m thi\u1ec3u hi\u1ec7n t\u01b0\u1ee3ng t\u1ea1o dimer ho\u1eb7c hairpin. Cu\u1ed1i c\u00f9ng, v\u1ecb tr\u00ed c\u1ee7a primer tr\u00ean gene c\u0169ng r\u1ea5t quan tr\u1ecdng, n\u00ean \u01b0u ti\u00ean c\u00e1c v\u00f9ng exon ho\u1eb7c intron \u1ed5n \u0111\u1ecbnh \u0111\u1ec3 tr\u00e1nh \u1ea3nh h\u01b0\u1edfng b\u1edfi c\u00e1c bi\u1ebfn th\u1ec3 splicing.<\/p>\n\n\n\n

5. V\u00ec sao kit Real-time PCR c\u1ee7a ABT l\u00e0 l\u1ef1a ch\u1ecdn t\u1ed1i \u01b0u?<\/strong><\/h2>\n\n\n\n

Kit Real-time PCR<\/a><\/strong> c\u1ee7a ABT \u0111\u01b0\u1ee3c \u0111\u00e1nh gi\u00e1 cao v\u1ec1 ch\u1ea5t l\u01b0\u1ee3ng nh\u1edd qu\u00e1 tr\u00ecnh s\u1ea3n xu\u1ea5t nghi\u00eam ng\u1eb7t v\u00e0 s\u1eed d\u1ee5ng c\u00e1c nguy\u00ean li\u1ec7u \u0111\u1ea1t chu\u1ea9n. C\u00e1c b\u1ed9 kit \u0111\u01b0\u1ee3c thi\u1ebft k\u1ebf t\u1ed1i \u01b0u, \u0111\u1ea3m b\u1ea3o hi\u1ec7u su\u1ea5t khu\u1ebfch \u0111\u1ea1i cao, \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u v\u01b0\u1ee3t tr\u1ed9i. M\u1ed7i s\u1ea3n ph\u1ea9m \u0111\u1ec1u tr\u1ea3i qua qu\u00e1 tr\u00ecnh ki\u1ec3m so\u00e1t ch\u1ea5t l\u01b0\u1ee3ng ch\u1eb7t ch\u1ebd \u0111\u1ec3 \u0111\u00e1p \u1ee9ng c\u00e1c ti\u00eau chu\u1ea9n qu\u1ed1c t\u1ebf, mang \u0111\u1ebfn k\u1ebft qu\u1ea3 ph\u00e2n t\u00edch ch\u00ednh x\u00e1c v\u00e0 \u0111\u00e1ng tin c\u1eady.<\/p>\n\n\n\n

ABT cung c\u1ea5p m\u1ed9t lo\u1ea1t c\u00e1c kit Real-time PCR \u0111\u00e1p \u1ee9ng \u0111a d\u1ea1ng nhu c\u1ea7u nghi\u00ean c\u1ee9u c\u1ee7a kh\u00e1ch h\u00e0ng v\u1edbi gi\u1ea3i ph\u00e1p ph\u00f9 h\u1ee3p, t\u1eeb vi\u1ec7c ph\u00e1t hi\u1ec7n c\u00e1c t\u00e1c nh\u00e2n truy\u1ec1n nhi\u1ec5m \u0111\u1ebfn \u0111\u1ecbnh l\u01b0\u1ee3ng n\u1ed3ng \u0111\u1ed9 m\u1ee5c ti\u00eau, \u0111\u00e1p \u1ee9ng d\u1ee5ng \u0111a d\u1ea1ng cho nhi\u1ec1u \u0111\u1ed1i t\u01b0\u1ee3ng x\u00e9t nghi\u1ec7m kh\u00e1c nhau nh\u01b0 ng\u01b0\u1eddi, th\u00fa y, th\u1ee7y s\u1ea3n, GMO\u2026 S\u1ef1 \u0111a d\u1ea1ng n\u00e0y cho ph\u00e9p kh\u00e1ch h\u00e0ng l\u1ef1a ch\u1ecdn s\u1ea3n ph\u1ea9m t\u1ed1i \u01b0u cho t\u1eebng nghi\u00ean c\u1ee9u c\u1ee5 th\u1ec3, t\u0103ng hi\u1ec7u qu\u1ea3 c\u00f4ng vi\u1ec7c.<\/p>\n\n\n\n

\u0110\u1ed9i ng\u0169 k\u1ef9 thu\u1eadt c\u1ee7a ABT lu\u00f4n s\u1eb5n s\u00e0ng h\u1ed7 tr\u1ee3 kh\u00e1ch h\u00e0ng trong su\u1ed1t qu\u00e1 tr\u00ecnh s\u1eed d\u1ee5ng s\u1ea3n ph\u1ea9m. T\u1eeb vi\u1ec7c t\u01b0 v\u1ea5n l\u1ef1a ch\u1ecdn s\u1ea3n ph\u1ea9m ph\u00f9 h\u1ee3p, thi\u1ebft k\u1ebf th\u00ed nghi\u1ec7m \u0111\u1ebfn gi\u1ea3i \u0111\u00e1p c\u00e1c th\u1eafc m\u1eafc k\u1ef9 thu\u1eadt, kh\u00e1ch h\u00e0ng s\u1ebd nh\u1eadn \u0111\u01b0\u1ee3c s\u1ef1 h\u1ed7 tr\u1ee3 t\u1eadn t\u00ecnh v\u00e0 chuy\u00ean nghi\u1ec7p. \u0110i\u1ec1u n\u00e0y gi\u00fap \u0111\u1ea3m b\u1ea3o qu\u00e1 tr\u00ecnh nghi\u00ean c\u1ee9u di\u1ec5n ra su\u00f4n s\u1ebb v\u00e0 hi\u1ec7u qu\u1ea3!<\/p>\n\n\n\n

6. T\u00e0i li\u1ec7u tham kh\u1ea3o<\/strong><\/h2>\n\n\n\n
    \n
  1. ABT, T. (2023, May 13). Tong hop kien thuc chi tiet ve ky thuat Real-time PCR<\/em>. Retrieved from ABT Biological Solutions Company Limited: https:\/\/abtvn.com\/real-time-pcr\/<\/li>\n\n\n\n
  2. Debode, F., Marien, A., Janssen, \u00c9., Bragard, C., & Berben, G. (2017). The influence of amplicon length on real-time PCR results. Base<\/em>.<\/li>\n\n\n\n
  3. Huong, H. H. (2003). Sinh hoc phan tu.<\/em> Ha Noi: Nha Xuat Ban Giao Duc.<\/li>\n\n\n\n
  4. IDT. (2024). What are the criteria for choosing a PCR amplicon length?<\/em> Retrieved from Integrated DNA Technologies: https:\/\/www.idtdna.com\/pages\/support\/faqs\/what-are-the-criteria-for-choosing-pcr-amplicon-length<\/li>\n\n\n\n
  5. Jain, A. (2020). Basic Techniques in Biochemistry, Microbiology and Molecular Biology.<\/em> USA: Springer Nature .<\/li>\n\n\n\n
  6. Lela Buckingham, Maribeth Flaws. (2007). Molecular diagnostics : fundamentals, methods, and clinical applications.<\/em> United States of America: F.A. Davis Company.<\/li>\n<\/ol>\n\n\n\n

    <\/p>","protected":false},"excerpt":{"rendered":"

    Real-time PCR (qPCR) cho ph\u00e9p x\u00e1c \u0111\u1ecbnh l\u01b0\u1ee3ng DNA sau m\u1ed7i chu k\u1ef3 th\u00f4ng qua thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang t\u1ea1o ra t\u00edn hi\u1ec7u hu\u1ef3nh quang t\u0103ng d\u1ea7n theo t\u1ef7 l\u1ec7 thu\u1eadn v\u1edbi s\u1ed1 l\u01b0\u1ee3ng s\u1ea3n ph\u1ea9m PCR (amplicon) \u0111\u01b0\u1ee3c t\u1ea1o ra (Lela Buckingham, Maribeth Flaws, 2007). M\u1ed9t trong nh\u1eefng y\u1ebfu t\u1ed1 quan tr\u1ecdng quy\u1ebft \u0111\u1ecbnh […]<\/p>","protected":false},"author":1,"featured_media":11937,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_uag_custom_page_level_css":"","site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"set","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center 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Team","author_link":"https:\/\/abtvn.com\/en\/author\/admin\/"},"uagb_comment_info":0,"uagb_excerpt":"Real-time PCR (qPCR) cho ph\u00e9p x\u00e1c \u0111\u1ecbnh l\u01b0\u1ee3ng DNA sau m\u1ed7i chu k\u1ef3 th\u00f4ng qua thu\u1ed1c nhu\u1ed9m hu\u1ef3nh quang t\u1ea1o ra t\u00edn hi\u1ec7u hu\u1ef3nh quang t\u0103ng d\u1ea7n theo t\u1ef7 l\u1ec7 thu\u1eadn v\u1edbi s\u1ed1 l\u01b0\u1ee3ng s\u1ea3n ph\u1ea9m PCR (amplicon) \u0111\u01b0\u1ee3c t\u1ea1o ra (Lela Buckingham, Maribeth Flaws, 2007). M\u1ed9t trong nh\u1eefng y\u1ebfu t\u1ed1 quan tr\u1ecdng quy\u1ebft \u0111\u1ecbnh…","_links":{"self":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/11743","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/comments?post=11743"}],"version-history":[{"count":6,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/11743\/revisions"}],"predecessor-version":[{"id":11994,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/posts\/11743\/revisions\/11994"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media\/11937"}],"wp:attachment":[{"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/media?parent=11743"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/categories?post=11743"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/abtvn.com\/en\/wp-json\/wp\/v2\/tags?post=11743"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}