Video: Nws raug nqi npaum li cas los hloov lub ligation?
2024 Tus sau: Miles Stephen | [email protected]. Kawg hloov kho: 2023-12-15 23:36
Kev hloov pauv . Ntxiv ntawm 1-5 µl ntawm ligation sib tov kom muaj peev xwm hlwb rau kev hloov pauv . Txuas ntxiv ligation nrog PEG ua rau poob rau hauv kev hloov pauv efficiency (Quick Ligation Kit). Electroporation raug pom zoo rau kev tsim loj (> 10,000 bp).
Ntawm no, ntau npaum li cas ligation khoom kuv xav tau rau kev hloov pauv?
Kev hloov pauv . Ntxiv ntawm 1-5 µl ntawm ligation sib tov kom muaj peev xwm hlwb rau kev hloov pauv . Txuas ntxiv ligation nrog PEG ua rau poob rau hauv kev hloov pauv efficiency (Quick Ligation Kit). Electroporation raug pom zoo rau kev tsim loj (> 10,000 bp).
Tom qab, lo lus nug yog, Ligation thiab transformation yog dab tsi? Hauv kev sim cloning ib txwm, cov kws tshawb nrhiav thawj zaug ntxig ib daim DNA, xws li cov noob, rau hauv ib lub voj voog ntawm DNA hu ua plasmid. Cov kauj ruam no siv txwv enzymes thiab DNA ligase thiab hu ua a ligation . Tom qab a ligation , cov kauj ruam tom ntej yog hloov DNA mus rau cov kab mob hauv cov txheej txheem hu ua kev hloov pauv.
Tom qab ntawd, ib tug kuj yuav nug, ntau npaum li cas DNA xav tau rau ib tug ligation?
Tag nrho cov concentration ntawm vector + ntxig yuav tsum nyob nruab nrab ntawm 1-10 Μg / ml kom ua tau zoo ligation . Vector: Ntxig molar piv ntawm 1: 1 thiab 1: 10 raug pom zoo (1: 3 yog qhov raug). Yog koj tsis paub meej txog koj li DNA concentrations, ua ntau yam ligations nrog sib txawv piv.
Koj tuaj yeem khaws cov tshuaj tiv thaiv ligation ntev npaum li cas?
Ligations tau yuav tsum tau ua nyob rau hauv chav tsev kub los yog txias (xav tias 12-16 ° C) hmo ntuj los yog txawm rau ob peb hnub, yog koj 'yuav tibneeg hu tauj coob. Koj ua tau kuj khaws ib lub ligation nyob rau hauv lub fridge thiab coj nws tawm tom qab mus txuas ntxiv ligating ntawm chav tsev kub rau li ntev raws li qhov tsim nyog.
Pom zoo:
Nws raug nqi npaum li cas los tsim ib qho chaw nyob hauv av?
Factory Built Storm Shelter Prices Premanufactured cua daj cua dub chaw yuav raug nqi tsawg li $3,300, suav nrog kev teeb tsa. Tus nqi nruab nrab ntawm 8 ft. los ntawm 10 ft. saum toj no-hauv av qauv yog nruab nrab ntawm $ 5,500 thiab $ 20,000
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