Chiŵerengero chochepa cha A260/A230 nthawi zambiri chimayamba chifukwa cha zonyansa zomwe zimakhala ndi kutalika kwa kutalika kwa 230nm.Tiyeni tiwone zomwe zonyansazi zikuphatikiza:

• Zowononga wamba	Mayamwidwe wavelength	Chiyerekezo zotsatira
  Mapuloteni	~ 230nm ndi 280nm	Kuchepetsa munthawi yomweyo A260/A 280ndi A260/A 280chiwerengero
  Mchere wa Guanidine	220-240 nm	Kuchepetsa A260/A 280chiŵerengero
  Phenol	~ 270nm	-
  Trizol	~ 230nm ndi 270nm	Kuchepetsa A260/A 280chiŵerengero
  EDTA	~ 230nm	Kuchepetsa A260/A 280chiŵerengero
  Ethanol	230-240 nm	Kuchepetsa A260/A 280chiŵerengero

 
 
 
Mayamwidwe wavelength ndi mtengo wosiyana wa zoipitsa wamba

Pkusokonezeka kwa minyewa
Kuwonongeka kwa mapuloteni kumatha kuonedwa ngati kuipitsidwa kofala kwambiri pakuchotsa kwa nucleic acid.Mapuloteni alipo pakati pa gawo lapamwamba lamadzimadzi ndi lapansiorganicgawo .Kuipitsa kudzachepetsa chiŵerengero cha A260/A280 ndi A260/A230 nthawi yomweyo, ndipo chiŵerengero cha A260/A230 chidzasintha moonekeratu kuposa chiŵerengero cha A260/A280.
Pamapeto pakesinthani mawuor qPCR zochita, zotsalira za mapuloteni zimatha kulepheretsa kapena kusokoneza ntchito ya enzyme.Njira yabwino yopewera kuipitsidwa kwa mapuloteni ndi kukumbukira mfundo yakuti "m'malo mocheperapo, pang'ono nthawi zambiri" pamene mukufuna mphamvu yamphamvu.

2. Kuwonongeka kwa Guanidinium
hydrochloride (GuHCl) ndi guanidine thiocyanate (GTC) ali ndi zotsatira za mapuloteni a denaturing, omwe amatha kuwononga ma cell membranes panthawi ya nucleic acid m'zigawo, zomwe zimapangitsa kuti mapuloteni azikhala ndi mpweya.Kutalika kwa mayamwidwe a GuHCl ndi GTC kuli pakati pa 220-240 nm, ndimchere wotsalira wa guanidinium udzachepetsa chiŵerengero cha A260/A230.Ngakhale mchere wotsalira wa guanidinium umachepetsa chiŵerengerocho,zotsatira za zoyeserera zapansi panthaka kwenikweni ndi zosafunika.

3. Kuwonongeka kwa Trizol
Chigawo chachikulu cha Trizol ndi phenol.Ntchito yayikulu ya phenol ndikutulutsa ma cell ndikutulutsa mapuloteni ndi nucleic acid mu cell.Ngakhale phenol imatha kusokoneza mapuloteni bwino, sichingalepheretse ntchito ya RNase.Choncho, 8 -hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, etc. amawonjezeredwa ku TRIzol kuti alepheretse RNase endogenous ndi exogenous RNase.
Pamene yopezera ma RNA, Trizol akhoza mofulumira lyse maselo ndi ziletsa nuclease anamasulidwa ku maselo, ndi otsala Trizol kwambiri kuchepetsa chiŵerengero cha A260/A230.
Processing njira: Pamene centrifuging, tisaiwale kuti phenol mu Trizol mosavuta sungunuka mu madzi gawo pansi chikhalidwe cha 4 ° ndi firiji.

4. Ethanol zotsalira
Ethanol imagwiritsidwa ntchito pomaliza kutulutsa DNA ndikusungunula ayoni amchere omwe amatha kumangika ku DNA.Mayamwidwe wavelength wapamwamba kwambirimayamwidwe pachimake chaMowa ulinso pa 230-240 nm, ameneidzachepetsanso chiŵerengero cha A260/A230.
Njira kupewa Mowa zotsalira akhoza kubwerezedwa kawiri pa elution komaliza, kuwomba mufume hoodkwa mphindi ziwiri kulola Mowa kuti asungunuke kwathunthu musanawonjezere chotchinga cha elution.
Komabe, ziyenera kudziwika kuti chiŵerengerochi ndi chilolezo chowunika cha khalidwe la RNA.Ngati ntchito zomwe tazitchulazi zikuyendetsedwa mosamalitsa, kupatuka pakati pa chiŵerengero ndi mlingo wokhazikika sikudzakhala ndi zotsatira zazikulu pakuyesera kumtunda.
Zogwirizana nazo:
Animal Total RNA Isolation zida
Plant Total RNA Isolation kit
Cell Total RNA Isolation zida
Plant Total RNA Isolation kit Plus