Video: Dab tsi feem ntau ua raws li DNA cloning vector?
2024 Tus sau: Miles Stephen | [email protected]. Kawg hloov kho: 2023-12-15 23:36
Muaj ntau hom cloning vectors , tab sis feem ntau feem ntau Cov khoom siv yog genetic engineering plasmids. Cloning Feem ntau ua ntej siv Escherichia coli, thiab cloning vectors nyob rau hauv E. coli muaj xws li plasmids, bacteriophages (xws li phage λ), cosmids, thiab cov kab mob uas muaj cov chromosomes (BACs).
Kuj nug, dab tsi yog vector hauv DNA cloning?
Hauv molecular cloning , a vector yog a DNA molecule siv los ua ib lub tsheb los tsim cov khoom siv txawv teb chaws mus rau lwm lub xovtooj ntawm tes, qhov twg nws tuaj yeem rov ua dua thiab / lossis qhia (piv txwv li, plasmid, cosmid, Lambda phages). A vector muaj txawv teb chaws DNA yog hu ua recombinant DNA.
Kuj Paub, ua li cas koj clone ib vector? Kev sim txheej txheem
- Khiav PCR thiab purify cov khoom PCR: Khiav PCR kom nthuav koj cov ntxig DNA.
- Digest koj DNA:
- Cais koj cov ntxig thiab vector los ntawm gel purification:
- Ligate koj qhov ntxig rau hauv koj lub vector:
- Kev hloov pauv:
- Isolate lub Finished Plasmid:
- Txheeb xyuas koj Plasmid los ntawm Sequencing:
Hauv qhov no, pBR322 ua haujlwm li cas cloning vector?
pBR322 Plasmid pBR322 yog ib qho ntawm thawj plasmids siv rau lub hom phiaj ntawm cloning . Nws muaj cov noob rau kev tiv thaiv tetracycline thiab ampicillin. Ntxig DNA ntawm qhov chaw txwv ua tau inactivate cov noob rau tetracycline (ib qho txiaj ntsig hu ua inactivation inactivation) lossis ampicillin kuj.
Cov txheej txheem twg tso cai rau ntau daim ntawv theej ntawm ntu tshwj xeeb ntawm DNA ua kom sai heev?
Polymerase chain reaction
Pom zoo:
Dab tsi yog 0.8 raws li ib feem feem?
Piv txwv tus nqi feem pua ntawm cov zauv feem 75% 0.75 3/4 80% 0.8 4/5 90% 0.9 9/10 99% 0.99 99/100
Dab tsi yog qhov pH ntau ntawm cov tshuab tsis siv neeg feem ntau?
Ntau lub tshuab buffering muaj nyob hauv lub cev uas pab tswj cov pH ntawm cov ntshav thiab lwm cov kua dej hauv qhov nqaim - nruab nrab ntawm pH 7.35 thiab 7.45. Ib qho tsis yog ib yam khoom uas tiv thaiv kev hloov pauv ntawm cov kua dej pH los ntawm kev nqus dej ntau dhau los yog hydroxyl ions
Yuav ua li cas koj sau ib feem raws li ib tug khoom ntawm ib tug tag nrho cov zauv thiab ib feem feem?
Cov cai los nrhiav cov khoom ntawm ib feem feem thiab tag nrho tus lej Peb xub sau tag nrho cov lej ua ib feem, piv txwv li, sau nws faib los ntawm ib qho; Piv txwv li: 7 yog sau ua 71. Peb mam li muab cov zauv. Peb muab cov denominators. Yog tias yuav tsum tau ua kom yooj yim, nws ua tiav thiab tom qab ntawd peb sau cov feem kawg
Dab tsi yog qhov tseeb vector thiab txheeb ze vector?
Thaum siv cov vector tiag tiag, tus kheej lub nkoj thiab lwm lub nkoj txav ntawm lawv qhov tseeb ceev thiab cov chav kawm. Cov vectors tseeb tuaj yeem paub qhov txawv ntawm cov hom phiaj txav mus thiab nyob ruaj khov. Cov txheeb ze vector pab nrhiav cov nkoj ntawm kev sib tsoo. Lub nkoj uas nws vector hla dhau ntawm tus kheej lub nkoj txoj haujlwm yog nyob rau hauv kev sib tsoo
Qhov twg ntawm cov ntsiab lus hauv qab no feem ntau pom nyob rau hauv feem ntau cov chiv?
Cov tshuaj chiv niaj hnub suav nrog ib lossis ntau tshaj ntawm peb lub ntsiab lus tseem ceeb tshaj plaws hauv cov zaub mov noj: nitrogen, phosphorus, thiab potassium. Qhov tseem ceeb thib ob yog cov ntsiab lus sulfur, magnesium, thiab calcium