Umlinganiselo ophantsi we-A260/A230 udla ngokubangelwa kukungacoceki ngowona mgangatho uphezulu wokufunxa wamaza kwi-230nm.Makhe sibone ukuba oku kungcola kubandakanya ntoni:

• Izingcolisi eziqhelekileyo	Ubude bamaza okufunxa	Umlinganiselo wesiphumo
  Iprotheni	~230nm kunye ne-280nm	Ukucuthwa ngaxeshanye kwe-A260/A 280kunye no-A260/A 280izilinganiso
  Ityuwa yeGuanidine	220-240 nm	Nciphisa i-A260/A 280umlinganiselo
  Phenol	~270nm	-
  Trizol	~230nm kunye ne-270nm	Nciphisa i-A260/A 280umlinganiselo
  EDTA	~230nm	Nciphisa i-A260/A 280umlinganiselo
  I-Ethanol	230-240 nm	Nciphisa i-A260/A 280umlinganiselo

 
 
 
Ubude bamaza okufunxa kunye nexabiso lokuchasana kwezingcolisi eziqhelekileyo

Pungcoliseko lwe-rotein
Ungcoliseko lweprotheyini lunokuthathwa njengolona ngcoliseko luqhelekileyo kwinkqubo yokutsalwa kwe-nucleic acid.Iprotheyini ikhona phakathi kwesigaba esiphezulu samanzi kunye nesisezantsieziphilayoisigaba .Ukungcola kuya kunciphisa umlinganiselo we-A260 / A280 kunye ne-A260 / A230 ngexesha elifanayo, kwaye umlinganiselo we-A260 / A230 uya kutshintsha ngokucacileyo kunomlinganiselo we-A260 / A280.
Ngexesha elizayoumva ukhupheloor Iimpendulo ze-qPCR, iiprotheyini eziseleyo zinokunqanda okanye ziphazamise umsebenzi we-enzyme.Eyona ndlela ilungileyo yokuphepha ukungcoliseka kweeprotheyini kukugcina engqondweni umgaqo othi "kunokuba ungaphantsi kokungaphezulu, isixa esincinci amaxesha amaninzi" xa unqwenela amandla amakhulu.

2. Ungcoliseko lwaseGuanidinium
i-hydrochloride (i-GuHCl) kunye ne-guanidine thiocyanate (GTC) inefuthe leeprotheni ze-denaturing, ezinokutshabalalisa ngokukhawuleza i-membrane zeseli ngexesha le-nucleic acid extraction process, ebangela i-protein denaturation kunye nemvula.Ubude be-absorption be-GuHCl kunye ne-GTC buphakathi kwe-220-240 nm, kwayeityuwa eseleyo ye-guanidinium iya kunciphisa umlinganiselo we-A260/A230.Nangona ityuwa eseleyo yeguanidinium iya kunciphisa umlinganiselo,impembelelo kwimifuniselo esezantsi ayinamsebenzi ngokwenene.

3. Usulelo lwe-Trizol
Inxalenye ephambili ye-Trizol yi-phenol.Umsebenzi oyintloko we-phenol kukukhupha iiseli kunye nokukhulula iiprotheni kunye nezinto ze-nucleic acid kwiiseli.Nangona i-phenol inokuthi isebenze ngokufanelekileyo iiprotheyini, ayikwazi ukuthintela ngokupheleleyo umsebenzi we-RNase.Ngoko ke, i-8 -hydroxyquinoline, i-guanidine isothiocyanate, i-β-mercaptoethanol, njl. yongezwa kwi-TRIzol ukuvimbela i-RNase engapheliyo kunye ne-exogenous.
Xa ukhupha i-RNA yeselula, i-Trizol inokukhupha ngokukhawuleza iiseli kwaye inqanda i-nuclease ekhutshwe kwiiseli, kwaye i-Trizol eseleyo iya kunciphisa kakhulu umlinganiselo we-A260 / A230.
Indlela yokucubungula: Xa i-centrifuging, kufuneka kuqatshelwe ukuba i-phenol e-Trizol i-soluble ngokulula kwisigaba samanzi phantsi kwemeko ye-4 ° kunye nokushisa kwegumbi.

4. Intsalela ye-Ethanol
I-Ethanol isetyenziselwa inkqubo yokugqibela yokunciphisa i-DNA ngelixa ukuchithwa i-ion yetyuwa enokuthi iboshwe kwi-DNA.Ubude be-absorption bophezuluincopho yokufunxaethanol kwakhona kwi 230-240 nm, leyoiya kunciphisa kwakhona umlinganiselo we-A260/A230.
Indlela yokuphepha i-ethanol residue inokuphinda iphindwe kabini ngexesha lokugqitywa kokugqibela, ukuvuthela kwiumsiimizuzu emibini ukuvumela i-ethanol ukuba ibe ngumphunga ngokupheleleyo phambi kokuba yongeze isithinteli solution.
Nangona kunjalo, kufuneka kwaziwe ukuba umlinganiselo sisalathiso sovavanyo lomgangatho we-RNA.Ukuba le misebenzi ikhankanywe ngasentla ilawulwa ngokungqongqo, ukutenxa phakathi komlinganiselo kunye noluhlu olusemgangathweni aluyi kuba nempembelelo enkulu kwiimvavanyo ezisezantsi.
Iimveliso ezinxulumeneyo:
Animal Total RNA Isolation kit
Plant Total RNA Isolation kit
ISeli iyonke ye-RNA Isolation kit
Plant Total RNA Isolation kit Plus