Qhov qis piv ntawm A260 / A230 feem ntau yog tshwm sim los ntawm impurities nrog qhov siab tshaj plaws nqus wavelength ntawm 230nm.Cia peb pom dab tsi cov impurities suav nrog:

• Cov pa phem	Kev nqus wavelength	Qhov cuam tshuam
  Protein	~ 230nm thiab 280nm	Ib txhij txo ntawm A260/A 280thiab A260/A 280piv
  Guanidine ntsev	220-240 nm	Txo cov A260/A 280piv
  Phenol	~ 270 nm	-
  Trizol	~ 230nm thiab 270nm	Txo cov A260/A 280piv
  EDTA	~ 230 nm	Txo cov A260/A 280piv
  Ethanol	230-240 nm	Txo cov A260/A 280piv

 
 
 
Absorption wavelength thiab qhov sib piv tus nqi ntawm cov pa phem

Protein paug
Protein pollution tuaj yeem suav tias yog cov pa phem tshaj plaws hauv cov txheej txheem nucleic acid extraction.Protein muaj nyob nruab nrab ntawm theem dej hauv dej thiab qis duaorganictheem.Cov pa phem yuav txo qhov piv ntawm A260 / A280 thiab A260 / A230 tib lub sijhawm, thiab qhov piv ntawm A260 / A230 yuav hloov ntau dua qhov piv ntawm A260 / A280.
Thaum lub sij hawm tom ntejrov qab transcriptionor qPCR cov tshuaj tiv thaiv, protein residues tuaj yeem cuam tshuam lossis cuam tshuam nrog kev ua haujlwm enzyme.Txoj hauv kev zoo tshaj plaws kom tsis txhob muaj cov kab mob protein ntau yog kom nco ntsoov lub hauv paus ntsiab lus ntawm "tsawg dua ntau dua, me me ntau zaus" thaum aspirating tus supernatant.

2. Guanidinium muaj kuab paug
hydrochloride (GuHCl) thiab guanidine thiocyanate (GTC) muaj cov nyhuv denaturing proteins, uas tuaj yeem ua kom puas cell membranes sai sai thaum lub sij hawm nucleic acid extraction txheej txheem, ua rau cov protein denaturation thiab nag lossis daus.Kev nqus wavelength ntawm GuHCl thiab GTC yog nyob nruab nrab ntawm 220-240 nm, thiabresidual guanidinium ntsev yuav txo qhov piv ntawm A260 / A230.Txawm hais tias qhov seem ntawm guanidinium ntsev yuav txo qhov piv,qhov cuam tshuam rau cov kev sim hauv qab no yog qhov tsis txaus ntseeg.

3. Trizol paug
Lub ntsiab tseem ceeb ntawm Trizol yog phenol.Lub luag haujlwm tseem ceeb ntawm phenol yog lyse hlwb thiab tso cov proteins thiab nucleic acid hauv cov hlwb.Txawm hais tias phenol tuaj yeem ua tau zoo denature proteins, nws tsis tuaj yeem cuam tshuam RNase kev ua haujlwm.Yog li ntawd, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, thiab lwm yam ntxiv rau TRIzol inhibit endogenous thiab exogenous RNase.
Thaum rho tawm cellular RNA, Trizol tuaj yeem ua rau lub hlwb sai sai thiab cuam tshuam cov nuclease tso tawm ntawm lub hlwb, thiab cov seem Trizol yuav txo qhov piv ntawm A260 / A230.
Txoj kev ua: Thaum centrifuging, nws yuav tsum tau muab sau tseg tias phenol nyob rau hauv Trizol yog yooj yim soluble nyob rau hauv cov dej theem nyob rau hauv lub mob ntawm 4 ° thiab chav tsev kub.

4. Ethanol residue
Ethanol yog siv nyob rau hauv cov txheej txheem kawg los ua kom cov DNA thaum yaj ntsev ions uas yuav raug khi rau DNA.Kev nqus wavelength ntawm qhov siab tshaj plawsabsorption ncov ntawmethanol kuj tseem nyob ntawm 230-240 nm, uastseem yuav txo qhov piv ntawm A260 / A230.
Txoj kev zam ntawm ethanol residue tuaj yeem rov ua dua ob zaug thaum lub sijhawm kawg elution, tshuab hauvfume hoodrau ob feeb kom tso cai rau ethanol kom tag nrho evaporate ua ntej ntxiv tsis rau elution.
Txawm li cas los xij, nws yuav tsum paub tias qhov sib piv tsuas yog qhov ntsuas ntsuas ntawm RNA zoo.Yog hais tias cov kev khiav hauj lwm saum toj no tau tswj nruj me ntsis, qhov sib txawv ntawm qhov sib piv thiab cov qauv ntau yam yuav tsis muaj kev cuam tshuam zoo rau kev sim hauv qab.
Yam khoom:
Animal Total RNA Isolation Kit
Plant Total RNA Isolation Kit
Cell Total RNA Isolation Kit
Plant Total RNA Isolation Kit Plus