Qhov Tseem Ceeb Qhov txawv - Microarray vs RNA Sequencing
Transcriptome sawv cev rau tag nrho cov ntsiab lus ntawm RNA tam sim no hauv ib lub xovtooj nrog rau mRNA, rRNA, tRNA, degraded RNA, thiab, tsis muaj degraded RNA. Profiling transcriptome yog txheej txheem tseem ceeb kom nkag siab txog kev nkag siab ntawm tes. Muaj ntau ntau txoj hauv kev rau transcriptome profileing. Microarray thiab RNA sequencing yog ob hom thev naus laus zis tsim los tshuaj xyuas cov ntawv sau tseg. Qhov sib txawv tseem ceeb ntawm microarray thiab RNA sequencing yog tias microarray yog raws li hybridization peev xwm ntawm predesigned labeled probes nrog lub hom phiaj cDNA sequences thaum RNA sequencing yog raws li qhov ncaj sequencing ntawm cDNA strands los ntawm advanced sequencing cov tswv yim xws li NGS. Microarray tau ua nrog kev paub ua ntej txog cov kab ke thiab RNA sequencing yog ua yam tsis muaj kev paub ua ntej txog sequences.
Microarray yog dab tsi?
Microarray yog ib txoj kev muaj zog, txhim khu kev qha thiab siab dhau los siv rau kev sau cov ntaub ntawv los ntawm cov kws tshawb fawb. Nws yog txoj hauv kev nrov tshaj plaws rau kev txheeb xyuas cov ntawv sau tseg. Nws yog tus nqi qis, uas nyob ntawm qhov kev sojntsuam hybridization.
Cov txheej txheem pib nrog kev rho tawm ntawm mRNA los ntawm cov qauv thiab kev tsim cov tsev qiv ntawv cDNA los ntawm tag nrho RNA. Tom qab ntawd nws yog tov nrog fluorescently sau npe predesigned probes ntawm ib tug khoom nto (spot matrix). Complementary sequences hybridize nrog cov ntawv sau npe sojntsuam hauv microarray. Tom qab ntawd microarray yog ntxuav thiab tshuaj ntsuam, thiab cov duab yog kom muaj nuj nqis. Sau cov ntaub ntawv yuav tsum tau soj ntsuam kom tau cov txheeb ze qhia profiles.
Qhov kev siv ntawm microarray probes yog suav tias yog proportional rau qhov muaj nuj nqis ntawm cov ntawv sau hauv cov qauv. Txawm li cas los xij, qhov tseeb ntawm cov txheej txheem yog nyob ntawm qhov tsim kev sojntsuam, kev paub ua ntej ntawm qhov sib lawv liag thiab affinity ntawm probes rau hybridization. Yog li microarray technology muaj kev txwv. Cov txheej txheem microarray tsis tuaj yeem ua tiav nrog cov ntawv sau qis. Nws ua tsis tau qhov sib txawv isoforms thiab txheeb xyuas cov kab mob sib txawv. Txij li txoj kev no yog nyob ntawm kev sib txuas ntawm kev sojntsuam, qee qhov teeb meem cuam tshuam nrog kev sib xyaw xws li hla-hybridization, nonspecific hybridization thiab lwm yam tshwm sim hauv cov txheej txheem microarray.
Daim duab 01: Microarray
RNA Sequencing yog dab tsi?
RNA phom sib luag (RNA seq) yog ib qho nyuam qhuav tsim tag nrho cov txheej txheem kev sib txuas lus. Nws yog txoj hauv kev nrawm thiab siab dhau ntawm kev sau cov ntaub ntawv profile. Nws ncaj qha ntsuas qhov kev qhia ntawm cov noob thiab ua rau kev tshawb nrhiav tob ntawm cov ntawv sau tseg. RNA seq tsis nyob ntawm qhov kev sojntsuam tau tsim los lossis kev paub ua ntej ntawm cov kab ke. Yog li ntawd, RNA seq txoj kev muaj siab rhiab heev thiab muaj peev xwm ntawm kev tshawb nrhiav cov noob tshiab thiab cov noob caj noob ces.
RNA sequencing txoj kev yog ua los ntawm ob peb kauj ruam. Tag nrho RNA ntawm lub xov tooj ntawm tes yuav tsum raug cais tawm thiab tawg. Tom qab ntawd, siv cov ntawv thim rov qab, lub tsev qiv ntawv cDNA yuav tsum tau npaj. Txhua cDNA strand yuav tsum tau ligated nrog adaptors. Tom qab ntawd cov tawg tawg yuav tsum tau nthuav dav thiab purified. Thaum kawg siv txoj kev NGS, kev sib sau ntawm cDNA yuav tsum tau ua.
Daim duab 02: RNA Sequencing
Qhov txawv ntawm Microarray thiab RNA sequencing yog dab tsi?
Microarray vs RNA Sequencing |
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| Microarray yog qhov muaj zog, txhim khu kev qha, siab dhau txoj kev. | RNA sequencing yog ib txoj hauv kev raug thiab siab dhau. |
| Qhov no yog tus nqi qis. | Nov yog txoj kev kim. |
| tshuaj ntsuam ntau tus qauv | |
| Qhov no ua kom yooj yim txheeb xyuas ntau tus qauv ib txhij. | Qhov no ua kom yooj yim txheeb xyuas ntau tus qauv. |
| Data Analysis | |
| Data analysis is complex. | Ntau cov ntaub ntawv raug tsim tawm hauv txoj kev no; li no, cov txheej txheem yog complex. |
| Kev Paub Txog Ua Ntej | |
| Txoj kev no yog ua raws li kev sojntsuam hybridization, yog li kev paub ua ntej ntawm cov kab ke hauv qhov xav tau. | Txoj kev no tsis yog nyob ntawm qhov kev paub ua ntej. |
| Txoj kev hloov pauv thiab keeb kwm tshiab | |
| Txoj kev no tsis tuaj yeem ntes cov qauv kev hloov pauv thiab cov noob tshiab. | Txoj kev no tuaj yeem txheeb xyuas cov qauv sib txawv xws li cov noob sib txuas, lwm qhov sib txuas, thiab cov noob tshiab. |
| xov xwm | |
| Qhov no tsis tuaj yeem kuaj pom qhov sib txawv ntawm kev qhia ntawm isoforms, yog li qhov no muaj qhov tsis txaus ntseeg. | Qhov no muaj qhov rhiab heev. |
| Txoj Kev | |
| Qhov no tsuas tuaj yeem ua rau muaj qhov sib txawv ntawm cov lus qhia. Qhov no tsis muab qhov tseeb ntawm cov noob qhia. | Nws muab cov lus qhia meej thiab txheeb ze. |
| Data Reanalysis | |
| Qhov no yuav tsum tau rov ua kom rov ua dua. | Cov ntaub ntawv sib txuas tuaj yeem rov ua dua. |
| Nyob rau cov neeg ua haujlwm tshwj xeeb thiab kev tsim kho vaj tse | |
| Kev tsim kho tshwj xeeb thiab cov neeg ua haujlwm tsis tas yuav tsum muaj rau microarray. | Tshwj xeeb infrastructure thiab cov neeg ua haujlwm xav tau los ntawm RNA sequencing. |
| Technical Issues | |
| Microarray txheej txheem muaj teeb meem xws li hla-hybridization, nonspecific hybridization, txwv tsis pub tus nqi ntawm tus neeg soj ntsuam, thiab lwm yam. | RNA seq cov txheej txheem zam cov teeb meem kev lag luam xws li hla-hybridization, nonspecific hybridization, txwv tsis pub kuaj tus nqi ntawm tus neeg soj ntsuam, thiab lwm yam. |
| Kev tsis sib haum | |
| Qhov no yog txoj kev tsis ncaj ncees vim nws nyob ntawm kev sib xyaw. | Kev tsis ncaj ncees yog qis piv rau microarray. |
Summary – Microarray vs RNA Sequencing
Microarray thiab RNA sequencing txoj kev yog siab dhau los ntawm platforms tsim rau transcriptome profiling. Ob txoj hauv kev tsim cov txiaj ntsig uas muaj feem cuam tshuam nrog cov noob qhia profiles. Txawm li cas los xij, RNA sequencing muaj qhov zoo dua microarray rau kev tshuaj ntsuam gene qhia. RNA sequencing yog ib txoj kev rhiab heev rau kev tshawb nrhiav cov ntaub ntawv tsis tshua muaj ntau dua li microarray. RNA sequencing kuj ua rau muaj kev sib txawv ntawm isoforms thiab kev txheeb xyuas cov noob variants. Txawm li cas los xij, microarray yog qhov kev xaiv zoo tshaj plaws ntawm cov kws tshawb fawb feem ntau vim tias RNA sequencing yog cov txheej txheem tshiab thiab kim nrog cov ntaub ntawv khaws cov kev sib tw thiab cov ntaub ntawv nyuaj.
