Video: Lub hom phiaj ntawm PCR khoom purification yog dab tsi?
2024 Tus sau: Miles Stephen | [email protected]. Kawg hloov kho: 2023-12-15 23:36
Kev huv huv DNA los ntawm a PCR Cov tshuaj tiv thaiv feem ntau tsim nyog rau kev siv qis, thiab pab tshem tawm cov enzymes, nucleotides, primers thiab tsis khoom. Kev lig kev cai, qhov no tau ua tiav los ntawm kev siv cov organic extraction, xws li phenol chloroform extraction, ua raws li ethanol nag lossis daus.
Tsis tas li cov lus nug yog, vim li cas nws thiaj li tsim nyog los ntxuav cov khoom PCR?
Koj yuav tsum tau purify PCR cov khoom txhawm rau tshem tawm cov tsis siv cov primers, nucleotides, thiab enzymes txhawm rau txhim kho txoj kev vam meej ntawm ligation, uas yuav tswj tau thiab ua raws cov txheej txheem. khoom ntawm PCR . Rau ua kom huv tus PCR cov khoom , qhov loj tsis suav nrog chromatography yog siv.
Tsis tas li ntawd, kuv yuav ua li cas tom qab PCR? Txhawm rau kom rov zoo, yaug lub khob retentate ob zaug nrog 10 mus rau 20 µL TE lossis dej. Qhov kawg PCR Cov tshuaj tiv thaiv tuaj yeem muaj txog li microgram ntawm amplified DNA uas tuaj yeem siv rau ntau yam kev siv tshuaj lom neeg molecular. Cov ntawv thov no yuav muaj ntau dua lossis tsawg dua rau cov khoom seem ntawm cov PCR tshuaj tiv thaiv sib tov.
Ib tug kuj yuav nug, koj ua li cas ntxuav cov khoom PCR?
Classical txoj kev siv los huv si nce PCR cov khoom Ua ntej ua ntu zus suav nrog gel electrophoresis, ethanol nag lossis daus, thiab kem chromatography. Txawm hais tias muaj txiaj ntsig zoo, cov txheej txheem no tuaj yeem ua tau sai sai thaum ua ntau yam qauv hauv qhov sib npaug thiab nthuav tawm qhov sib txawv ntawm cov qauv poob.
Vim li cas thiaj yuav tsum tau ntxuav DNA?
Peb huv si nce DNA los ntawm cov tshuaj aqueous kom tshem tawm cov ntsev tsis muaj ntsev, enzymes lossis lwm yam khoom uas tuaj yeem cuam tshuam rau cov ntawv thov hauv qab. Piv txwv xws li cov tshuaj tiv thaiv PCR huv si nce, huv si tom qab kev txwv digests thiab huv si nce ntawm genomic los yog plasmid DNA kis nrog cellular proteins / khib nyiab.
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