Vim li cas thiaj li yuav tsum tau ua kom huv PCR cov khoom ua ntej sequencing?

2024 Tus sau: Stanley Ellington | [email protected]. Kawg hloov kho: 2023-12-16 00:17

Kev txheeb xyuas qhov xav tau khoom yog qhov tseem ceeb , uas suav nrog kev lees paub qhov tsis muaj qhov tsis tshwj xeeb cov khoom thiab primer dimers. huv -up ntawm cov tshuaj tiv thaiv sib tov kuj tsim nyog tshem tawm cov primers thiab dNTPs uas tsis tuaj yeem cuam tshuam nrog cov kev tshwm sim tom ntej thiab ua rau muaj qhov tsis tau nyeem.

Tsis tas li ntawd, 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.

Ib tug kuj nug, koj puas yuav tsum ntxuav PCR cov khoom ua ntej kev txwv endonuclease digestion? Feem ntau ntawm txwv enzymes yog active nyob rau hauv PCR buffers. Rau kev siv cloning, ua kom huv ntawm PCR cov khoom ua ntej kev zom zaub mov yog tsim nyog tshem tawm cov active thermophilic DNA polymerase tam sim no nyob rau hauv lub PCR sib tov. DNA polymerases tuaj yeem hloov kho qhov kawg ntawm DNA cleaved thiab txo cov txiaj ntsig ntawm ligation.

Tib neeg kuj nug, koj npaj PCR cov khoom lag luam li cas rau kev sib txuas?

Direct Sequencing ntawm PCR cov khoom

1. Xyuas kom tseeb tias koj tau nthuav dav cov kab ncaj nraim.
2. Koj yuav tsum tshem tawm tag nrho cov seem PCR primers thiab unincorporated nucleotides.
3. Yog tias PCR primers tseem yuav yog cov txheej txheem ua ntu zus, xyuas kom lawv phim peb cov xwm txheej.
4. Tsis txhob tshaj qhov ua piv txwv!

Koj puas tuaj yeem ua raws li PCR khoom?

Ncaj qha Sequencing ntawm PCR cov khoom . Sequencing siv xwb ib primer es tsis txhob ntawm ob siv nyob rau hauv PCR . Yog koj ua tsis tshem tawm ob qho tib si primers, koj yuav qib ob ua ntu zus superimposed rau ib leeg uas nyeem tsis tau. Nws yog OK siv a PCR primer rau ua ke tsuav nws phim peb tej yam kev mob.