Qhov txawv ntawm nanopore thiab illumina sequencing yog nanopore sequencing yog ib tiam thib peb-tiam sequencing txheej txheem uas siv ib tug nanopore los ntes cov sequence ntawm DNA molecule, thaum illumina sequencing yog ib tug thib ob-tiam sequencing txheej txheem uas siv reversible. dye terminators thev naus laus zis los txheeb xyuas qhov sib lawv liag ntawm DNA molecule.
DNA sequencing yog qhov kev txiav txim siab ntawm cov nucleotide meej lossis lub hauv paus ntawm DNA molecule. Muaj ntau ntau txoj hauv kev los txiav txim siab nucleic acid sequences uas ua rau kom muaj kev tshawb fawb txog kev lom neeg thiab kev kho mob. Ib qho ntawm thawj DNA sequencing cov txheej txheem (Sanger sequencing) yog tsim los ntawm Frederick Sanger nyob rau hauv 1975 los ntawm kev siv primer extension lub tswv yim ntawm MRC Center, Cambridge, UK. Niaj hnub no, feem ntau ntawm cov txheej txheem DNA nrawm nrawm yog nyob rau tiam thib ob (tiam tom ntej) thiab tiam thib peb DNA sequencing pawg. Nanopore thiab illumina sequencing yog ob yam tshiab DNA technologies.
Nanopore Sequencing yog dab tsi?
Nanopore sequencing yog txheej txheem thib peb-tiam sequencing uas siv cov protein nanopore los kuaj cov nucleic acid sib lawv liag ntawm DNA molecule. Hauv nanopore sequencing, DNA dhau los ntawm nanopore hloov nws tam sim no. Qhov kev hloov pauv no nyob ntawm cov duab, qhov loj, thiab qhov ntev ntawm DNA ib ntus. Cov teeb liab tshwm sim tau txiav txim siab kom tau txais cov DNA tshwj xeeb lossis RNA ib ntus. Txoj kev no tsis tas yuav hloov kho nucleotides, thiab nws ua tau raws sijhawm.
Daim duab 01: Nanopore Sequencing
Oxford Nanopore Technologies yog lub tuam txhab nrov uas tsim cov khoom siv ntau yam nanopore sequencing. Feem ntau ntawm Oxford Nanopore sequencing li muaj cov hlwb khiav. Cov ndlwg ntawm tes no muaj ib tug xov tooj ntawm me me nanopores uas yog embedded nyob rau hauv electro resistant membrane. Txhua nanopore sib raug rau nws tus kheej electrode. Qhov no electrode txuas mus rau ib channel thiab ib tug sensor nti. Cov electrode no ntsuas cov hluav taws xob tam sim no ntws los ntawm nanopore. Thaum ib lub molecule hla dhau nanopore, nws tam sim no hloov los yog cuam tshuam. Tsis tas li ntawd, qhov kev cuam tshuam no ua rau muaj tus cwj pwm squiggle. Qhov squiggle no yog tom qab txiav txim siab los txiav txim siab DNA lossis RNA ib ntus hauv lub sijhawm tiag tiag.
Illumina Sequencing yog dab tsi?
Illumina sequencing yog cov txheej txheem thib ob thib ob uas siv cov thev naus laus zis thim rov qab dye terminators thev naus laus zis txhawm rau txheeb xyuas cov kab ke ntawm DNA molecules. Solexa lub tuam txhab, tam sim no yog ib feem ntawm lub tuam txhab Illumina, tau tsim muaj nyob rau hauv 1998. Lub tuam txhab no tau tsim cov txheej txheem sequencing raws li cov txheej txheem thim rov qab dye terminators technology thiab engineeringed polymerases.
Daim duab 02: Illumina Sequencing
Nyob rau hauv illumina sequencing txoj kev, tus qauv yog thawj cleaved rau hauv luv luv seem. Yog li ntawd, nyob rau hauv illumina sequencing, 100-150bp luv luv nyeem los yog fragments yog tsim thaum pib. Cov fragments no ces ligated rau generic adapters thiab annealed rau ib tug swb. PCR tau ua tiav los nthuav tawm txhua ntu. Qhov no tsim ib qho chaw nrog ntau cov ntawv luam ntawm tib qhov tawg. Tom qab ntawd, lawv tau muab cais ua ib leeg-stranded thiab raug rau sequencing. Cov kab sib txuas ua ke muaj cov tshuaj fluorescently sau npe nucleotides, DNA polymerase, thiab cov terminator. Vim yog lub terminator, tsuas yog ib lub hauv paus ntxiv ib zaug. Txhua lub voj voog terminator raug tshem tawm, thiab nws tso cai rau qhov sib ntxiv ntawm lub hauv paus tom ntej mus rau qhov chaw. Tsis tas li ntawd, raws li cov teeb liab fluorescent, lub khoos phis tawj kuaj pom lub hauv paus ntxiv hauv txhua lub voj voog. Illumina sequencing thev naus laus zis tsim cov kab ke hauv 4 txog 56 teev.
Dab tsi zoo sib xws ntawm Nanopore thiab Illumina Sequencing?
- Nanopore thiab illumina sequencing yog ob txoj kev ua ke.
- Ob leeg yog txoj hauv kev nrawm nrawm thiab tshiab.
- Lawv siv los kuaj DNA thiab RNA ib ntus.
- Ob leeg muaj qhov raug siab.
Qhov txawv ntawm Nanopore thiab Illumina Sequencing yog dab tsi?
Nanopore sequencing yog txheej txheem thib peb-tiam sequencing uas siv nanopore los kuaj xyuas qhov sib lawv liag ntawm DNA molecules. Nyob rau hauv sib piv, illumina sequencing yog ib tug thib ob-tiam sequencing txheej txheem uas siv reversible dye terminators thev naus laus zis los xyuas qhov sib lawv liag ntawm DNA molecules. o, qhov no yog qhov sib txawv tseem ceeb ntawm nanopore thiab illumina sequencing. Ntxiv mus, nanopore sequencing muaj 92-97% raug, thaum illumina sequencing muaj 99% raug.
Cov infographic hauv qab no teev qhov sib txawv ntawm nanopore thiab illumina sequencing nyob rau hauv daim ntawv tabular.
Summary – Nanopore vs Illumina Sequencing
Cov txheej txheem kev sib tw siab muaj xws li tiam thib ob (luam luv) thiab tiam thib peb (ntev-nyeem) txoj hauv kev. Nanopore thiab illumina sequencing yog ob lub DNA tshiab thev naus laus zis uas muaj nyob rau tiam thib peb thiab thib ob tiam (tom ntej tiam) DNA sequencing pawg. Nanopore sequencing siv lub nanopore los xyuas qhov sib lawv liag ntawm DNA molecules. Ntawm qhov tod tes, illumina sequencing siv reversible dye terminators thev naus laus zis los txheeb xyuas qhov sib lawv liag ntawm DNA molecules. Yog li, qhov no yog cov ntsiab lus ntawm qhov sib txawv ntawm nanopore thiab illumina sequencing.
