Isilinganiso esiphansi se-A260/A230 sivamise ukudalwa ukungcola okunomkhawulo wamaza wamaza wamaza wokumunca ku-230nm.Ake sibone ukuthi lokhu kungcola kufaka phakathi:

• Ukungcola okuvamile	Ubude begagasi bokumuncwa	Umphumela wesilinganiso
  Amaprotheni	~230nm kanye ne-280nm	Ukwehliswa ngesikhathi esisodwa kwe-A260/A 280kanye no-A260/A 280izilinganiso
  Usawoti we-Guanidine	220-240 nm	Nciphisa i-A260/A 280isilinganiso
  I-Phenol	~270nm	-
  I-Trizol	~230nm kanye ne-270nm	Nciphisa i-A260/A 280isilinganiso
  I-EDTA	~230nm	Nciphisa i-A260/A 280isilinganiso
  I-Ethanol	230-240 nm	Nciphisa i-A260/A 280isilinganiso

 
 
 
Ubude begagasi bokumuncwa kanye nevelu yokuqhathanisa yezinto ezingcolile ezivamile

Pukutheleleka kwe-rotein
Ukungcola kwamaprotheni kungathathwa njengokungcoliswa okuvame kakhulu enqubweni yokukhipha i-nucleic acid.Amaprotheni akhona phakathi kwesigaba se-aqueous esiphezulu nesingaphansieziphilayoisigaba .Ukungcola kuzonciphisa isilinganiso se-A260/A280 ne-A260/A230 ngesikhathi esifanayo, futhi isilinganiso se-A260/A230 sizoshintsha ngokusobala kakhulu kunesilinganiso se-A260/A280.
Ngesikhathi esilandelayoukuhlehla okulotshiweyoor Ukusabela kwe-qPCR, izinsalela zamaprotheni zingavimbela noma ziphazamise ukusebenza kwe-enzyme.Indlela engcono kakhulu yokugwema ukungcoliswa kwamaprotheni iwukukhumbula isimiso esithi “kunokuncane kunokuningi, inani elincane izikhathi eziningi” lapho ulangazelela amandla angaphezu kwavamile.

2. Ukungcola kwe-Guanidinium
i-hydrochloride (GuHCl) kanye ne-guanidine thiocyanate (GTC) zinomphumela wamaprotheni e-denaturing, angabhubhisa ngokushesha ulwelwesi lwamangqamuzana ngesikhathi senqubo yokukhipha i-nucleic acid, okubangela ukuguqulwa kwamaprotheni kanye nemvula.Ubude begagasi bokumuncwa be-GuHCl ne-GTC buphakathi kuka-220-240 nm, kanyeusawoti osele we-guanidinium uzonciphisa isilinganiso se-A260/A230.Nakuba usawoti osele we-guanidinium uzonciphisa isilinganiso,umthelela ocwaningweni olwenzeka phansi empeleni awunaki.

3. Ukungcoliswa kwe-Trizol
Ingxenye eyinhloko ye-Trizol yi-phenol.Umsebenzi oyinhloko we-phenol ukuhlanganisa amaseli nokukhulula amaprotheni nezinto ze-nucleic acid kumaseli.Nakuba i-phenol ingakwazi ukuguqula amaprotheni ngokuphumelelayo, ayikwazi ukuvimbela ngokuphelele umsebenzi we-RNase.Ngakho-ke, i-8 -hydroxyquinoline, i-guanidine isothiocyanate, i-β-mercaptoethanol, njll. yengezwa ku-TRIzol ukuvimbela i-RNa engapheli kanye ne-exogenous.
Lapho ikhipha i-RNA yeselula, i-Trizol ingakwazi ukuhlanganisa amaseli ngokushesha futhi ivimbele i-nuclease ekhishwe kumaseli, futhi i-Trizol esele izonciphisa kakhulu isilinganiso se-A260/A230.
Indlela yokucubungula: Uma i-centrifuging, kufanele kuqashelwe ukuthi i-phenol ku-Trizol incibilika kalula esigabeni samanzi ngaphansi kwesimo se-4 ° kanye negumbi lokushisa.

4. Izinsalela ze-Ethanol
I-Ethanol isetshenziswa enqubweni yokugcina ukuze kuncishiswe i-DNA ngenkathi kuchithwa ama-ion kasawoti angase aboshelwe ku-DNA.Ubude begagasi bokumuncwa obuphakeme kakhuluukumuncwa isiqongo ofI-ethanol nayo iku-230-240 nm, okuyintoizophinde yehlise isilinganiso se-A260/A230.
Indlela yokugwema insalela ye-ethanol ingaphindaphindwa kabili ngesikhathi sokukhishwa kokugcina, ivuthela ku-i-fume hoodimizuzu emibili ukuvumela i-ethanol ukuba ihwamuke ngokugcwele ngaphambi kokwengeza isivali ukuze i-lution.
Nokho, kufanele kwaziwe ukuthi isilinganiso siyinkomba yokuhlola yekhwalithi ye-RNA kuphela.Uma imisebenzi eshiwo ngenhla ilawulwa ngokuqinile, ukuchezuka phakathi kwesilinganiso nobubanzi obujwayelekile ngeke kube nomthelela omkhulu ekuhlolweni komfula.
Imikhiqizo Ehlobene:
Ikhithi yokuhlukanisa i-RNA yezilwane
Ikhithi yokuhlukanisa i-RNA Yezitshalo
Ikhithi Yokuhlukanisa Yeseli Yengqikithi ye-RNA
Plant Total RNA Isolation kit Plus