Gibson cloning ua haujlwm li cas?
Gibson cloning ua haujlwm li cas?

Video: Gibson cloning ua haujlwm li cas?

Video: Gibson cloning ua haujlwm li cas?
Video: Дизайн праймера и сборка фрагментов с использованием сборки NEBuilder HiFi DNA или сборки Gibson 2024, Kaum ib hlis
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Cov Gibson Cloning Master Mix muaj peb qhov sib txawv enzymes hauv ib qho tsis muaj. Txhua enzyme muaj qhov tshwj xeeb thiab tshwj xeeb ua haujlwm rau cov tshuaj tiv thaiv: T5 Exonuclease - tsim ib leeg-strand DNA 3' overhangs los ntawm zom rov qab los ntawm DNA 5' kawg. Ntxiv DNA fragments tuaj yeem txuas ntxiv rau ib leeg.

Xav txog qhov no, Gibson ua haujlwm li cas?

Cov Gibson Assembly® txoj kev yog txheej txheem cloning uas tso cai rau lub cloning ntawm ob los yog ntau tshaj fragments yam tsis tas yuav muaj kev txwv enzyme digestion los yog tshaj qhov chaw txwv. Hloov chaw, cov neeg siv-txhais lus sib tshooj kawg tau muab tso rau hauv cov khoom tawg kom tso cai rau kev sib koom ua ke ntawm cov seem uas nyob ib sab.

Ib yam li ntawd, hauv Gibson Assembly Master Mix yog dab tsi? Cov Gibson Cloning Master Mix muaj peb lub enzymes sib txawv hauv ib qho tsis muaj. Txhua enzyme muaj qhov tshwj xeeb thiab tshwj xeeb ua haujlwm rau cov tshuaj tiv thaiv: T5 Exonuclease - tsim ib leeg-strand DNA 3' overhangs los ntawm zom rov qab los ntawm DNA 5' kawg. Ntxiv DNA fragments tuaj yeem txuas ntxiv rau ib leeg.

Hauv qhov no, Golden Gate sib dhos ua haujlwm li cas?

Golden Rooj vag cloning lub Golden Gate los ua ke yog ib txoj kev molecular cloning uas tso cai rau tus kws tshawb fawb los sib sau ua ke ntau yam DNA tawg rau hauv ib daim siv hom IIs txwv enzymes thiab T4 DNA ligase. Qhov no sib dhos yog ua hauv vitro.

Gateway cloning ua haujlwm li cas?

Cov GATEWAY Cloning Technology yog raws li lub site-specific recombination system siv los ntawm phage l los integrate nws DNA nyob rau hauv lub E. AttL thiab attR chaw nyob ib ncig ntawm lub inserted phage DNA recombine site-tshwj xeeb thaum lub sij hawm excision kev tshwm sim los hloov lub attP site nyob rau hauv phage l thiab cov attB site hauv E.

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