{"id":13788,"date":"2025-10-28T15:09:40","date_gmt":"2025-10-28T07:39:40","guid":{"rendered":"https:\/\/abtvn.com\/?p=13788"},"modified":"2025-12-29T12:23:38","modified_gmt":"2025-12-29T04:53:38","slug":"dpcr-dua-tren-chip-va-tren-giot-hay-nhu-tuong","status":"publish","type":"post","link":"https:\/\/abtvn.com\/en\/dpcr-dua-tren-chip-va-tren-giot-hay-nhu-tuong\/","title":{"rendered":"H\u01b0\u1edbng d\u1eabn th\u1ef1c hi\u1ec7n PCR k\u1ef9 thu\u1eadt s\u1ed1 (dPCR) d\u1ef1a tr\u00ean chip v\u00e0 tr\u00ean gi\u1ecdt hay nh\u0169 t\u01b0\u01a1ng<\/strong>"},"content":{"rendered":"

PCR k\u1ef9 thu\u1eadt s\u1ed1 (Digital PCR – dPCR)<\/a><\/strong> l\u00e0 k\u1ef9 thu\u1eadt \u0111\u1ecbnh l\u01b0\u1ee3ng tuy\u1ec7t \u0111\u1ed1i axit nucleic trong n\u1ec1n m\u1eabu ph\u1ee9c t\u1ea1p v\u1edbi \u0111\u1ed9 nh\u1ea1y v\u00e0 \u0111\u1ed9 \u0111\u1eb7c hi\u1ec7u cao. dPCR th\u01b0\u1eddng \u0111\u01b0\u1ee3c ph\u00e2n lo\u1ea1i th\u00e0nh PCR k\u1ef9 thu\u1eadt s\u1ed1 d\u1ef1a tr\u00ean chip (cdPCR)<\/strong> and PCR k\u1ef9 thu\u1eadt s\u1ed1 d\u1ef1a tr\u00ean gi\u1ecdt\/nh\u0169 t\u01b0\u01a1ng (ddPCR\/edPCR)<\/strong>. B\u00e0i vi\u1ebft n\u00e0y h\u01b0\u1edbng d\u1eabn th\u1ef1c hi\u1ec7n cdPCR tr\u00ean h\u1ec7 th\u1ed1ng DcentriGene 160 (Bioer) v\u00e0 ddPCR tr\u00ean h\u1ec7 th\u1ed1ng QX200™ Droplet Digital™ PCR (Bio-rad).<\/p>\n\n\n\n

1. H\u01b0\u1edbng d\u1eabn th\u1ef1c hi\u1ec7n cdPCR tr\u00ean h\u1ec7 th\u1ed1ng DcentriGene 160 (Bioer)<\/strong><\/h2>\n\n\n\n

H\u1ec7 th\u1ed1ng ph\u00e2n t\u00edch PCR k\u1ef9 thu\u1eadt s\u1ed1 d\u1ef1a tr\u00ean chip (cdPCR) l\u00e0 ph\u01b0\u01a1ng ph\u00e1p ph\u00e2n chia ph\u1ea3n \u1ee9ng v\u00e0o c\u00e1c khoang ph\u1ea3n \u1ee9ng c\u00f3 th\u1ec3 t\u00edch si\u00eau nh\u1ecf nh\u1edd b\u1ed9 t\u1ea1o vi gi\u1ecdt (MicroDrop Generator). H\u1ed7n h\u1ee3p ph\u1ea3n \u1ee9ng cdPCR \u0111\u01b0\u1ee3c ph\u00e2n t\u00e1ch th\u00e0nh 10.000 \u0111\u1ebfn 45.000 khoang tr\u00ean m\u1ed9t chip. Sau \u0111\u00f3, chip \u0111\u01b0\u1ee3c \u0111\u01b0a v\u00e0o h\u1ec7 th\u1ed1ng khu\u1ebfch \u0111\u1ea1i b\u1eb1ng m\u00e1y lu\u00e2n nhi\u1ec7t (dPCR Thermal Cycler). T\u00edn hi\u1ec7u hu\u1ef3nh quang t\u1eeb t\u1ea5t c\u1ea3 c\u00e1c khoang tr\u00ean chip \u0111\u01b0\u1ee3c ghi nh\u1eadn b\u1eb1ng m\u1ed9t b\u1ed9 \u0111\u1ecdc vi chip (MicroChip Reader) trang b\u1ecb camera c\u00f4ng su\u1ea5t cao v\u00e0 b\u1ed9 l\u1ecdc hu\u1ef3nh quang. Ph\u1ea7n m\u1ec1m ph\u00e2n t\u00edch s\u1ebd t\u00ednh to\u00e1n ch\u00ednh x\u00e1c s\u1ed1 b\u1ea3n sao c\u1ee7a tr\u00ecnh t\u1ef1 m\u1ee5c ti\u00eau m\u00e0 kh\u00f4ng c\u1ea7n x\u00e2y d\u1ef1ng \u0111\u01b0\u1eddng chu\u1ea9n.<\/p>\n\n\n\n

\"H\u00ecnh
Figure 1. <\/strong>H\u1ec7 th\u1ed1ng DcentriGene 160 (Bioer)<\/em><\/figcaption><\/figure>\n\n\n\n

B\u1ed9 t\u1ea1o vi gi\u1ecdt (H\u00ecnh 1A)<\/em><\/strong> c\u00f3 c\u00f4ng ngh\u1ec7 t\u1ea1o gi\u1ecdt d\u1ef1a tr\u00ean ly t\u00e2m \u0111\u1ec3 ph\u00e2n t\u00e1ch m\u1eabu th\u00e0nh 25.000\u201332.000 vi gi\u1ecdt \u0111\u1ed3ng nh\u1ea5t, t\u1ed1i \u01b0u h\u00f3a vi\u1ec7c s\u1eed d\u1ee5ng m\u1eabu. M\u00e1y lu\u00e2n nhi\u1ec7t dPCR (H\u00ecnh 1B)<\/em><\/strong> c\u00f3 b\u1ed1n d\u1ea3i ki\u1ec3m so\u00e1t nhi\u1ec7t \u0111\u1ed9 \u0111\u1ed9c l\u1eadp, cho ph\u00e9p th\u1ef1c hi\u1ec7n PCR \u0111\u1ed3ng th\u1eddi tr\u00ean b\u1ed1n m\u1eabu. B\u1ed9 \u0111\u1ecdc vi chip (H\u00ecnh 1C) <\/em><\/strong>ghi nh\u1eadn t\u00edn hi\u1ec7u hu\u1ef3nh quang \u0111\u1ec3 ph\u00e2n bi\u1ec7t vi gi\u1ecdt d\u01b0\u01a1ng t\u00ednh ch\u1ee9a gen \u0111\u00edch \u0111\u00e3 khu\u1ebfch \u0111\u1ea1i v\u00e0 vi gi\u1ecdt \u00e2m t\u00ednh. Ph\u1ea7n m\u1ec1m ph\u00e2n t\u00edch \u00e1p d\u1ee5ng ph\u00e2n b\u1ed1 Poisson \u0111\u1ec3 \u0111\u1ecbnh l\u01b0\u1ee3ng ch\u00ednh x\u00e1c axit nucleic m\u1ee5c ti\u00eau.<\/p>\n\n\n\n

Quy tr\u00ecnh v\u1eadn h\u00e0nh h\u1ec7 th\u1ed1ng DcentriGene 160 c\u1ee7a Bioer nh\u01b0 sau:<\/em><\/strong><\/p>\n\n\n\n

B\u01b0\u1edbc 1: N\u1ea1p m\u1eabu v\u00e0o chip t\u1ea1o vi gi\u1ecdt<\/em><\/strong><\/p>\n\n\n\n

M\u1eabu \u0111\u00e3 t\u00e1ch chi\u1ebft v\u00e0 tinh s\u1ea1ch \u0111\u01b0\u1ee3c n\u1ea1p v\u00e0o MicroDrop Chip (Th\u1ec3 t\u00edch 15 \u00b5L). Sau \u0111\u00f3, g\u1eafn c\u00e1c chip ch\u1ee9a m\u1eabu v\u00e0o thi\u1ebft b\u1ecb t\u1ea1o gi\u1ecdt (H\u00ecnh 1A)<\/em><\/strong>, thi\u1ebft b\u1ecb n\u00e0y c\u00f3 th\u1ec3 t\u1ea1o ra 25.000 – 32.000 gi\u1ecdt v\u1edbi 16 MicroDrop Chip t\u01b0\u01a1ng \u1ee9ng v\u1edbi s\u1ed1 m\u1eabu t\u1ed1i \u0111a c\u00f3 th\u1ec3 ch\u1ea1y l\u00e0 16 m\u1eabu. Khi thi\u1ebft b\u1ecb quay ly t\u00e2m, c\u00e1c chip xoay l\u00e0m ch\u1ea5t l\u1ecfng ch\u1ea3y t\u1eeb gi\u1ebfng v\u00e0o c\u00e1c k\u00eanh t\u1ea1o gi\u1ecdt. 16 gi\u1ecdt m\u1eabu tr\u1ea3i \u0111\u1ec1u tr\u00ean chip trong kho\u1ea3ng 3 ph\u00fat v\u1edbi t\u1ec9 l\u1ec7 s\u1eed d\u1ee5ng m\u1eabu tr\u00ean 95%.<\/p>\n\n\n\n

\"H\u00ecnh
Figure 2: <\/strong>C\u00e1c b\u01b0\u1edbc n\u1ea1p m\u1eabu v\u00e0o chip t\u1ea1o vi gi\u1ecdt<\/em><\/figcaption><\/figure>\n\n\n\n

B\u01b0\u1edbc 2: Khu\u1ebfch \u0111\u1ea1i vi gi\u1ecdt<\/em><\/strong><\/p>\n\n\n\n

C\u00e1c chip ch\u1ee9a vi gi\u1ecdt \u0111\u01b0\u1ee3c chuy\u1ec3n sang thi\u1ebft b\u1ecb PCR \u0111\u1ec3 khu\u1ebfch \u0111\u1ea1i. Chu tr\u00ecnh nhi\u1ec7t (t\u0103ng – gi\u1ea3m nhi\u1ec7t \u0111\u1ed9) \u0111\u01b0\u1ee3c c\u00e0i \u0111\u1eb7t ri\u00eang cho t\u1eebng chip \u0111\u1ed9c l\u1eadp. Thi\u1ebft b\u1ecb c\u00f3 4 v\u00f9ng \u0111i\u1ec1u khi\u1ec3n nhi\u1ec7t \u0111\u1ed9 \u0111\u1ed9c l\u1eadp t\u1ea1o s\u1ef1 \u0111\u1ed3ng \u0111\u1ec1u v\u1ec1 m\u1eb7t nhi\u1ec7t \u0111\u1ed9 gi\u1eefa c\u00e1c m\u1eabu. T\u1ed5ng th\u1eddi gian khu\u1ebfch \u0111\u1ea1i c\u00e1c vi gi\u1ecdt kho\u1ea3ng 70 ph\u00fat cho 1-16 m\u1eabu v\u1edbi \u0111\u1ed9 ch\u00ednh x\u00e1c \u22640.1\u2103.<\/p>\n\n\n\n

\"H\u00ecnh
H\u00ecnh 3: <\/strong>C\u00e1c chip \u0111\u1eb7t v\u00e0o thi\u1ebft b\u1ecb PCR<\/em><\/figcaption><\/figure>\n\n\n\n

B\u01b0\u1edbc 3: \u0110\u1ecdc t\u00edn hi\u1ec7u ghi nh\u1eadn<\/em><\/strong><\/p>\n\n\n\n

Sau qu\u00e1 tr\u00ecnh khu\u1ebfch \u0111\u1ea1i, c\u00e1c MicroDrop Chip \u0111\u01b0\u1ee3c chuy\u1ec3n sang thi\u1ebft b\u1ecb \u0111\u1ecdc t\u00edn hi\u1ec7u \u0111\u1ec3 \u0111\u1ecbnh t\u00ednh v\u00e0 \u0111\u1ecbnh l\u01b0\u1ee3ng m\u1eabu. Thi\u1ebft b\u1ecb \u0111\u1ecdc t\u00edn hi\u1ec7u v\u1edbi 6 k\u00eanh m\u00e0u t\u01b0\u01a1ng \u1ee9ng l\u00e0 FAM\/HEX\/ROX\/CY5\/CY5.5\/CY3, c\u00e1c b\u01b0\u1edbc s\u00f3ng hu\u1ef3nh quang l\u1ea7n l\u01b0\u1ee3t \u0111\u01b0\u1ee3c ph\u00e1t ra v\u00e0 thu nh\u1eadn t\u00edn hi\u1ec7u tr\u00ean m\u1ed7i chip d\u1ef1a tr\u00ean c\u00f4ng ngh\u1ec7 CCD \u0111\u1ec3 \u0111\u1ecbnh l\u01b0\u1ee3ng axit nucleic trong t\u1eebng gi\u1ecdt. Th\u1eddi gian ph\u00e2n t\u00edch 16 m\u1eabu 6 k\u00eanh trong kho\u1ea3ng 25 ph\u00fat v\u00e0 th\u1eddi gian 1 k\u00eanh 1 chip d\u01b0\u1edbi 10 gi\u00e2y.<\/p>\n\n\n\n

\"H\u00ecnh
H\u00ecnh 4: <\/strong>C\u00e1c chip sau khi khu\u1ebfch \u0111\u1ea1i \u0111\u01b0\u1ee3c \u0111\u1eb7t v\u00e0o m\u00e1y \u0111\u1ecdc<\/em><\/figcaption><\/figure>\n\n\n\n

B\u01b0\u1edbc 4: Ph\u00e2n t\u00edch d\u1eef li\u1ec7u b\u1eb1ng ph\u1ea7n m\u1ec1m<\/em><\/strong><\/p>\n\n\n\n

Sau khi \u0111\u1ecdc t\u00edn hi\u1ec7u hu\u1ef3nh quang, bi\u1ec3u \u0111\u1ed3 t\u00edn hi\u1ec7u \u0111\u01b0\u1ee3c m\u00e1y xu\u1ea5t ra tr\u00ean ph\u1ea7n m\u1ec1m cho ph\u00e9p quan s\u00e1t k\u1ebft qu\u1ea3 theo k\u00eanh m\u00e0u l\u1ef1a ch\u1ecdn. Ph\u00e2n t\u00edch th\u1ed1ng k\u00ea Poisson cho th\u1ea5y ph\u1ea7n tr\u00ean l\u00e0 t\u00edn hi\u1ec7u c\u00e1c gi\u1ecdt d\u01b0\u01a1ng t\u00ednh v\u00e0 d\u01b0\u1edbi l\u00e0 c\u00e1c gi\u1ecdt \u00e2m t\u00ednh. K\u1ebft qu\u1ea3 c\u0169ng \u0111\u01b0\u1ee3c xu\u1ea5t d\u01b0\u1edbi d\u1ea1ng b\u1ea3ng \u0111\u1ecbnh l\u01b0\u1ee3ng tuy\u1ec7t \u0111\u1ed1i c\u1ee7a t\u1eebng m\u1eabu gi\u00fap ng\u01b0\u1eddi s\u1eed d\u1ee5ng d\u1ec5 d\u00e0ng \u0111\u1ecdc v\u00e0 ph\u00e2n t\u00edch k\u1ebft qu\u1ea3.<\/p>\n\n\n\n

\"H\u00ecnh
H\u00ecnh 5: <\/strong>K\u1ebft qu\u1ea3 ph\u00e2n t\u00edch d\u1eef li\u1ec7u b\u1eb1ng ph\u1ea7n m\u1ec1m<\/em><\/figcaption><\/figure>\n\n\n\n

2. H\u01b0\u1edbng d\u1eabn th\u1ef1c hi\u1ec7n ddPCR tr\u00ean h\u1ec7 th\u1ed1ng QX200 Droplet Digital PCR (Bio-Rad)<\/strong><\/h2>\n\n\n\n

H\u1ec7 th\u1ed1ng QX200 Droplet Digital PCR (ddPCR) c\u1ee7a Bio-Rad bao g\u1ed3m thi\u1ebft b\u1ecb QX200 Droplet Generator d\u00f9ng \u0111\u1ec3 ph\u00e2n chia m\u1eabu t\u1eeb 20 \u00b5L th\u00e0nh 20.000 gi\u1ecdt c\u00f3 k\u00edch th\u01b0\u1edbc si\u00eau nh\u1ecf. M\u00e1y PCR khu\u1ebfch \u0111\u1ea1i v\u1edbi ch\u1ea5t ph\u00e1t quang EvaGreen ho\u1eb7c \u0111\u1ea7u d\u00f2 th\u1ee7y ph\u00e2n TaqMan Probe. M\u00e1y QX200 Droplet Reader \u0111\u1ecdc t\u00edn hi\u1ec7u t\u1eeb c\u00e1c gi\u1ecdt \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh m\u1eabu d\u01b0\u01a1ng t\u00ednh v\u00e0 \u00e2m t\u00ednh nh\u1eb1m \u0111\u1ecbnh l\u01b0\u1ee3ng tuy\u1ec7t \u0111\u1ed1i l\u01b0\u1ee3ng DNA m\u1ee5c ti\u00eau.<\/p>\n\n\n\n

\"H\u00ecnh
H\u00ecnh 6: <\/strong>Quy tr\u00ecnh chung th\u1ef1c hi\u1ec7n PCR k\u1ef9 thu\u1eadt s\u1ed1 vi gi\u1ecdt tr\u00ean h\u1ec7 th\u1ed1ng QX200 Droplet Digital PCR (Bio-Rad)<\/em><\/figcaption><\/figure>\n\n\n\n

V\u1ec1 c\u01a1 b\u1ea3n, c\u00e1c gi\u1ecdt khu\u1ebfch \u0111\u1ea1i ch\u1ee9a h\u00f3a ch\u1ea5t v\u00e0 quy tr\u00ecnh l\u00e0m vi\u1ec7c t\u01b0\u01a1ng t\u1ef1 nh\u01b0 x\u00e9t nghi\u1ec7m qPCR d\u1ef1a tr\u00ean \u0111\u1ea7u d\u00f2 TaqMan. Sau khu\u1ebfch \u0111\u1ea1i, m\u1ed7i gi\u1ecdt \u0111\u01b0\u1ee3c \u0111\u1ecdc v\u00e0 ph\u00e2n t\u00edch \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh t\u1ef7 l\u1ec7 c\u00e1c gi\u1ecdt PCR d\u01b0\u01a1ng t\u00ednh ho\u1eb7c \u00e2m t\u00ednh. Sau \u0111\u00f3, d\u1eef li\u1ec7u n\u00e0y \u0111\u01b0\u1ee3c ph\u00e2n t\u00edch b\u1eb1ng th\u1ed1ng k\u00ea Poisson \u0111\u1ec3 x\u00e1c \u0111\u1ecbnh n\u1ed3ng \u0111\u1ed9 khu\u00f4n m\u1eabu DNA m\u1ee5c ti\u00eau trong m\u1eabu ban \u0111\u1ea7u.<\/p>\n\n\n\n

Quy tr\u00ecnh th\u1ef1c hi\u1ec7n tr\u00ean h\u1ec7 th\u1ed1ng QX200 Droplet Digital PCR c\u1ee7a Bio-Rad \u0111\u01b0\u1ee3c th\u1ef1c hi\u1ec7n nh\u01b0 sau:<\/strong><\/p>\n\n\n\n

B\u01b0\u1edbc 1: Chu\u1ea9n b\u1ecb m\u1eabu v\u00e0 ph\u1ea3n \u1ee9ng<\/em><\/strong><\/p>\n\n\n\n

Ch\u1ea5t l\u01b0\u1ee3ng c\u1ee7a qu\u00e1 tr\u00ecnh t\u00e1ch chi\u1ebft axit nucleic c\u00f3 th\u1ec3 \u1ea3nh h\u01b0\u1edfng \u0111\u1ebfn k\u1ebft qu\u1ea3 ddPCR. Do \u0111\u00f3, c\u1ea7n s\u1eed d\u1ee5ng ph\u01b0\u01a1ng ph\u00e1p t\u1ed1i \u01b0u \u0111\u1ec3 t\u00e1ch chi\u1ebft DNA ho\u1eb7c RNA t\u1eeb m\u1eabu ban \u0111\u1ea7u, \u0111\u1ea3m b\u1ea3o r\u1eb1ng m\u1eabu kh\u00f4ng b\u1ecb ph\u00e2n h\u1ee7y. \u0110\u1ed3ng th\u1eddi, qu\u00e1 tr\u00ecnh tinh s\u1ea1ch axit nucleic n\u00ean lo\u1ea1i b\u1ecf t\u1ed1i \u0111a c\u00e1c ch\u1ea5t \u1ee9c ch\u1ebf PCR. Trong tr\u01b0\u1eddng h\u1ee3p kh\u00f4ng th\u1ec3 lo\u1ea1i b\u1ecf ho\u00e0n to\u00e0n, c\u00f3 th\u1ec3 pha lo\u00e3ng m\u1eabu theo t\u1ef7 l\u1ec7 1:10 tr\u01b0\u1edbc khi ti\u1ebfn h\u00e0nh ph\u1ea3n \u1ee9ng.<\/p>\n\n\n\n

Th\u00e0nh ph\u1ea7n ph\u1ea3n \u1ee9ng ddPCR v\u1ec1 c\u01a1 b\u1ea3n t\u01b0\u01a1ng t\u1ef1 nh\u01b0 ph\u1ea3n \u1ee9ng PCR th\u00f4ng th\u01b0\u1eddng, bao g\u1ed3m m\u1ed3i, \u0111\u1ea7u d\u00f2 ho\u1eb7c ch\u1ea5t ph\u00e1t quang, m\u1eabu DNA v\u00e0 h\u1ed7n h\u1ee3p master mix ch\u1ee9a dNTP, Mg2+<\/sup>, enzyme DNA polymerase\u2026v\u1edbi t\u1ed5ng th\u1ec3 t\u00edch cu\u1ed1i c\u00f9ng c\u1ee7a ph\u1ea3n \u1ee9ng l\u00e0 20 \u03bcL.<\/p>\n\n\n\n

L\u01b0u \u00fd: Thu\u1ed1c th\u1eed ddPCR tr\u00ean h\u1ec7 th\u1ed1ng c\u1ee7a Bio-Rad l\u00e0 s\u1ea3n ph\u1ea9m \u0111\u1ed9c quy\u1ec1n \u0111\u01b0\u1ee3c ph\u00e1t tri\u1ec3n d\u00e0nh ri\u00eang cho vi\u1ec7c t\u1ea1o gi\u1ecdt tr\u00ean h\u1ec7 th\u1ed1ng c\u1ee7a Bio-Rad.<\/em><\/p>\n\n\n\n

B\u01b0\u1edbc 2: T\u1ea1o vi gi\u1ecdt<\/em><\/strong><\/p>\n\n\n\n

S\u1eed d\u1ee5ng pipet h\u00fat 20 \u00b5L th\u1ec3 t\u00edch h\u1ed7n h\u1ee3p \u0111\u00e3 ph\u1ed1i tr\u1ed9n \u1edf tr\u00ean v\u00e0o c\u00e1c h\u00e0ng gi\u1eefa c\u1ee7a DG8TM<\/sup> cartridge ngay v\u1ecb tr\u00ed \u0111\u00e1nh d\u1ea5u \u201csample\u201d.<\/p>\n\n\n\n

H\u00fat 70 \u00b5L d\u1ea7u chuy\u1ec3n v\u00e0o v\u1ecb tr\u00ed \u201cOil\u201d v\u00e0 g\u1eafn 1 mi\u1ebfng \u0111\u1ec7m \u1edf ph\u00eda tr\u00ean tr\u01b0\u1edbc khi \u0111\u1eb7t cartridge v\u00e0o m\u00e1y t\u1ea1o gi\u1ecdt. M\u1ed9t ph\u1ea3n \u1ee9ng 20\u00a0\u03bcL s\u1ebd \u0111\u01b0\u1ee3c ph\u00e2n chia ng\u1eabu nhi\u00ean th\u00e0nh 20.000 vi gi\u1ecdt c\u00f3 th\u1ec3 t\u00edch si\u00eau nh\u1ecf trong kho\u1ea3ng 2.5 ph\u00fat cho 8 m\u1eabu. C\u00e1c ph\u00e2n t\u1eed DNA \u0111\u01b0\u1ee3c ph\u00e2n b\u1ed1 ng\u1eabu nhi\u00ean v\u00e0o m\u1ed7i vi gi\u1ecdt v\u00e0 tr\u1edf th\u00e0nh 1 ph\u1ea3n \u1ee9ng ri\u00eang bi\u1ec7t. <\/p>\n\n\n\n

\"H\u00ecnh
Figure 7.<\/strong> C\u00e1c b\u01b0\u1edbc t\u1ea1o vi gi\u1ecdt b\u1eb1ng h\u1ec7 th\u1ed1ng QX200 droplet generator<\/em><\/figcaption><\/figure>\n\n\n\n

L\u01b0u \u00fd: \u0110\u1ec3 \u0111\u1ea3m b\u1ea3o h\u00fat ch\u00ednh x\u00e1c 20 \u00b5L h\u1ed7n h\u1ee3p chuy\u1ec3n v\u00e0o cartridge, n\u00ean chu\u1ea9n b\u1ecb ph\u1ea3n \u1ee9ng ban \u0111\u1ea7u l\u1edbn h\u01a1n 20 \u00b5L m\u1ed9t ch\u00fat (22-25 \u00b5L). C\u00e1c h\u1ed7n h\u1ee3p ph\u1ea3n \u1ee9ng ph\u1ea3i \u0111\u01b0\u1ee3c tr\u1ed9n \u0111\u1ec1u tr\u01b0\u1edbc khi chuy\u1ec3n v\u00e0o cartridge 8 gi\u1ebfng, trong qu\u00e1 tr\u00ecnh th\u1ef1c hi\u1ec7n c\u1ea7n n\u1ea1p c\u1ea3 c\u00e1c gi\u1ebfng ch\u01b0a s\u1eed d\u1ee5ng b\u1eb1ng dung d\u1ecbch \u0111\u1ec7m.<\/em><\/p>\n\n\n\n

B\u01b0\u1edbc 3: Ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i c\u00e1c vi gi\u1ecdt<\/em><\/strong><\/p>\n\n\n\n

Sau khi t\u1ea1o th\u00e0nh vi gi\u1ecdt, ti\u1ebfn h\u00e0nh h\u00fat nh\u1eb9 nh\u00e0ng c\u00e1c gi\u1ecdt t\u1ea1o th\u00e0nh chuy\u1ec3n v\u00e0o \u0111\u0129a PCR 96 gi\u1ebfng. Sealing \u0111\u0129a PCR 96 gi\u1ebfng v\u00e0 \u0111\u1eb7t v\u00e0o m\u00e1y PCR khu\u1ebfch \u0111\u1ea1i theo ch\u01b0\u01a1ng tr\u00ecnh c\u00e0i \u0111\u1eb7t nh\u01b0 m\u1ed9t ph\u1ea3n \u1ee9ng PCR truy\u1ec1n th\u1ed1ng.<\/p>\n\n\n\n

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Figure 8. <\/strong>Ph\u1ea3n \u1ee9ng khu\u1ebfch \u0111\u1ea1i c\u00e1c vi gi\u1ecdt v\u00e0 c\u00e0i \u0111\u1eb7t th\u00f4ng s\u1ed1 tr\u00ean ph\u1ea7n m\u1ec1m QuantaSoftTM<\/sup><\/em><\/figcaption><\/figure>\n\n\n\n

B\u01b0\u1edbc 4: \u0110\u1ecdc t\u00edn hi\u1ec7u trong c\u00e1c gi\u1ecdt<\/em><\/strong><\/p>\n\n\n\n

Sau khi khu\u1ebfch \u0111\u1ea1i axit nucleic m\u1ee5c ti\u00eau trong t\u1eebng gi\u1ecdt, ti\u1ebfn h\u00e0nh \u0111\u1eb7t \u0111\u0129a PCR v\u00e0o m\u00e1y \u0111\u1ecdc QX200 droplet reader. T\u1eebng vi gi\u1ecdt \u0111\u01b0\u1ee3c t\u00e1ch ra ri\u00eang l\u1ebb v\u00e0 l\u1ea7n l\u01b0\u1ee3t \u0111i qua c\u00e1c \u0111\u1ea7u \u0111\u1ecdc t\u00edn hi\u1ec7u hu\u1ef3nh quang b\u1eb1ng m\u00e1y \u0111\u1ecdc gi\u1ecdt. T\u00edn hi\u1ec7u hu\u1ef3nh quang \u0111\u01b0\u1ee3c \u0111o v\u00e0 ghi nh\u1eadn cho t\u1eebng gi\u1ecdt ri\u00eang l\u1ebb.<\/p>\n\n\n\n

M\u00e1y \u0111\u1ecdc gi\u1ecdt v\u00e0 ph\u1ea7n m\u1ec1m QuantaSoft \u0111\u1ebfm c\u00e1c gi\u1ecdt PCR d\u01b0\u01a1ng t\u00ednh v\u00e0 PCR \u00e2m t\u00ednh \u0111\u1ec3 cung c\u1ea5p s\u1ed1 l\u01b0\u1ee3ng tuy\u1ec7t \u0111\u1ed1i c\u1ee7a DNA m\u1ee5c ti\u00eau c\u00f3 trong t\u1eebng gi\u1ecdt. Th\u00f4ng th\u01b0\u1eddng, c\u00e1c gi\u1ecdt d\u01b0\u01a1ng t\u00ednh ch\u1ee9a \u00edt nh\u1ea5t m\u1ed9t b\u1ea3n sao c\u1ee7a ph\u00e2n t\u1eed DNA m\u1ee5c ti\u00eau s\u1ebd bi\u1ec3u hi\u1ec7n hu\u1ef3nh quang t\u0103ng l\u00ean so v\u1edbi c\u00e1c gi\u1ecdt \u00e2m t\u00ednh.<\/p>\n\n\n\n

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H\u00ecnh 9:<\/strong> \u0110\u1ecdc t\u00edn hi\u1ec7u trong t\u1eebng gi\u1ecdt b\u1eb1ng QX200 Droplet Reader<\/em><\/figcaption><\/figure>\n\n\n\n

B\u01b0\u1edbc 5: Ph\u00e2n t\u00edch d\u1eef li\u1ec7u<\/em><\/strong><\/p>\n\n\n\n

D\u1eef li\u1ec7u tr\u00ean t\u1eebng gi\u1ebfng ph\u1ea3i \u0111\u00e1p \u1ee9ng ch\u1ea5t l\u01b0\u1ee3ng nh\u1ea5t \u0111\u1ecbnh tr\u01b0\u1edbc khi ph\u1ea7n m\u1ec1m QuantSoft t\u1ef1 \u0111\u1ed9ng t\u00ednh to\u00e1n ng\u01b0\u1ee1ng m\u00e0 tr\u00ean \u0111\u00f3 c\u00e1c gi\u1ecdt \u0111\u01b0\u1ee3c coi l\u00e0 d\u01b0\u01a1ng t\u00ednh. Ng\u01b0\u1ee1ng c\u00f3 th\u1ec3 \u0111\u01b0\u1ee3c \u0111i\u1ec1u ch\u1ec9nh th\u1ee7 c\u00f4ng theo t\u1eebng gi\u1ebfng ho\u1eb7c tr\u00ean to\u00e0n b\u1ed9 \u0111\u0129a.<\/p>\n\n\n\n

K\u1ebft qu\u1ea3 c\u00f3 th\u1ec3 xem d\u01b0\u1edbi d\u1ea1ng bi\u1ec3u \u0111\u1ed3 1-D, trong \u0111\u00f3:<\/strong><\/p>\n\n\n\n