Cov txheej txheem:
Video: Yuav ua li cas koj ntxig cov noob rau hauv plasmid?
2024 Tus sau: Stanley Ellington | [email protected]. Kawg hloov kho: 2024-01-18 08:18
Cov kauj ruam yooj yim yog:
- Txiav qhib lub plasmid thiab "paste" hauv lub noob . Cov txheej txheem no tso siab rau kev txwv enzymes (uas txiav DNA) thiab DNA ligase (uas koom nrog DNA).
- Ntxig tus plasmid ua kab mob.
- Loj hlob ntau plasmid - nqa cov kab mob thiab siv lawv ua "factories" los ua cov protein.
Tsis tas li ntawd, yuav ua li cas cov noob tau muab tso rau hauv plasmid GCSE?
nws yog muab tso rau hauv plasmid siv ligase enzymes. tus plasmid mus rau hauv kab mob cell. cov kab mob transgenic reproduces, tshwm sim hauv tsheej lab ntawm cov kab mob zoo tib yam uas tsim cov tshuaj insulin rau tib neeg.
Tsis tas li ntawd, ua li cas koj Subclone ib tug noob? Cov kauj ruam yooj yim rau Subcloning Koj tso tawm thiab ntxuav koj cov ntxig los ntawm niam txiv vector, ligate no ntxig rau hauv qhov chaw npaj vector, hloov qhov ligation cov tshuaj tiv thaiv rau hauv cov kab mob muaj peev xwm. Tom qab ntawd koj tshuaj ntsuam cov hlwb hloov pauv rau qhov ntxig.
Ua raws li qhov no hauv kev xav, dab tsi yog 4 kauj ruam ntawm noob cloning?
Nyob rau hauv classical txwv enzyme digestion thiab ligation cloning raws tu qauv, cloning ntawm tej DNA fragment yeej tseem ceeb muaj plaub kauj ruam:
- Kev cais tawm ntawm DNA ntawm kev txaus siab (lossis lub hom phiaj DNA),
- kev ligation,
- transfection (los yog transformation), thiab.
- ib txoj kev tshuaj ntsuam/xaiv.
Yuav ua li cas amplify plasmid?
Kev sim txheej txheem
- Khiav PCR thiab ntxuav cov khoom PCR: Khiav PCR kom nthuav dav 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:
- Tshawb xyuas koj Plasmid los ntawm Sequencing:
Pom zoo:
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