Iyo yakaderera reshiyo yeA260/A230 inowanzokonzerwa nekusachena ine yakanyanya kunyura wavelength pa230nm.Ngationei kuti tsvina iyi inosanganisirei:

• Kazhinji zvinosvibisa	Absorption wavelength	Ratio effect
  Mapuroteni	~ 230nm uye 280nm	Panguva imwe chete kuderedzwa kweA260/A 280uye A260/A 280ratios
  Guanidine munyu	220-240 nm	Kuderedza A260/A 280ratio
  Phenol	~270nm	-
  Trizol	~ 230nm uye 270nm	Kuderedza A260/A 280ratio
  EDTA	~230nm	Kuderedza A260/A 280ratio
  Ethanol	230-240 nm	Kuderedza A260/A 280ratio

 
 
 
Absorption wavelength uye kukosha kwekusiyanisa kwezvakajairwa zvinosvibisa

Pkusvibiswa kwerotein
Kusvibiswa kweprotein kunogona kutorwa seyakanyanya kusvibiswa muiyo nucleic acid yekubvisa maitiro.Mapuroteni aripo pakati pechikamu chepamusoro chemvura uye chepasiorganicphase .Kusvibiswa kuchaderedza chiyero cheA260 / A280 uye A260 / A230 panguva imwe chete, uye chiyero cheA260 / A230 chichachinja zviri pachena kupfuura chiyero cheA260 / A280.
Munguva inoteverareverse transcriptionor qPCR maitiro, mapuroteni akasara anogona kutadzisa kana kukanganisa basa re enzyme.Nzira yakanakisa yekudzivisa kusvibiswa kweprotein ndeyekurangarira musimboti wekuti "panzvimbo shoma pane yakawanda, chiyero chidiki kakawanda" paunenge uchishuvira simba repamusoro.

2. Guanidinium kusvibiswa
hydrochloride (GuHCl) uye guanidine thiocyanate (GTC) ine mhedzisiro yedenaturing mapuroteni, anogona kukurumidza kuparadza maseru membranes panguva yeiyo nucleic acid yekubvisa maitiro, zvichikonzera mapuroteni denaturation uye kunaya.Kutora wavelength yeGuHCl neGTC iri pakati pe220-240 nm, uyeyakasara guanidinium munyu inoderedza reshiyo yeA260/A230.Kunyangwe iyo yakasara guanidinium munyu inoderedza reshiyo,mabatirwo anoitwa zviyedzo zvepasi haakoshesi.

3. Trizol kusvibiswa
Chinhu chikuru cheTrizol ndicho phenol.Basa guru re phenol ndere lyse masero uye kusunungura mapuroteni uye nucleic acid zvinhu mumasero.Kunyangwe phenol inogona kunyatso dhiza mapuroteni, haigone kumisa zvachose RNase basa.Nokudaro, 8 -hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, nezvimwewo zvinowedzerwa kuTRIzol kuti inhibit endogenous uye exogenous RNase.
Paunenge uchibvisa cellular RNA, Trizol inogona kukurumidza kumisa maseru uye inhibit nuclease inoburitswa kubva mumasero, uye yakasara Trizol inoderedza zvakanyanya chiyero cheA260 / A230.
Nzira yekugadzirisa: Kana centrifuging, inofanira kuonekwa kuti phenol muTrizol inonyungudika nyore nyore muchikamu chemvura pasi pemamiriro e4 ° uye tembiricha yekamuri.

4. Ethanol yakasara
Ethanol inoshandiswa mukugadzirisa kwekupedzisira kupinza DNA panguva yekunyungudutsa maion emunyu anogona kusungirirwa kuDNA.Kutorwa kwewavelength kwepamusoro-soroabsorption peak ofethanol zvakare iri pa230-240 nm, iyoichaderedzawo chiyero cheA260/A230.
Iyo nzira yekudzivisa ethanol yakasara inogona kudzokororwa kaviri panguva yekupedzisira elution, ichivhuvhuta mukatifume hoodkwemaminetsi maviri kubvumira ethanol kuti inyunguduke zvakazara isati yawedzera buffer yekubuda.
Nekudaro, zvinofanirwa kuzivikanwa kuti reshiyo ingori yekuongorora indekisi yeRNA mhando.Kana mashandiro ataurwa pamusoro akanyatsogadziriswa, kutsauka pakati pereshiyo uye mwero weyero hakuzove nekukanganisa kukuru pazviyedzo zvepasi.
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