DNase I (Rnase Mahala)(5u/ul)
Inombolo yekati: HC4007A
I-DNase I (i-Deoxyribonuclease I) yi-endodeoxyribonuclease ekwazi ukwetyisa i-DNA enemisonto emibini.Iqaphela kwaye iqhawule iibhondi ze-phosphodiester ukuvelisa i-monodeoxynucleotides okanye i-oligodeoxynucleotides enye okanye kabini kunye namaqela e-phosphate kwi-5'-terminal kunye ne-hydroxyl kwi-3'-terminal.Umsebenzi we-DNase I uxhomekeke kwiCa2+kwaye inokuthi isebenze ngee-ion zetsimbi ze-divalent ezifana noMn2+kunye noZn2+.5 mM Ca2+ikhusela i-enzyme kwi-hydrolysis.Phambi kukaMg2+, le enzyme inokuzibona ize icande kuyo nayiphi na indawo kuwo nawuphi na umcu weDNA.Phambi kukaMnu2+, imisonto ephindwe kabini ye-DNA inokubonwa ngaxeshanye kwaye icandeke phantse kwindawo enye ukuze yenze amaqhekeza e-DNA asekupheleni okanye amaqhekeza ancangathi ancangathi e-DNA ane-1-2 yeenucleotide eziphumayo.
Amacandelo
| Igama | 0.1KU | 1KU | 5 KU | 50 KU |
| DNase I, RNase-free | 20μL | 200μL | 1mL | 10 ml |
| 10×DNase I Buffer | 1mL | 1mL | 5 × 1mL | 5 × 10 mL |
Iimeko zokugcina
-25℃~-15℃ yokugcina;Uthutho phantsi kweepakethe zeqhwa.
Imiyalelo
1. Lungiselela isisombululo sokuphendula kwityhubhu ye-RNase-free ngokomlinganiselo odweliswe ngezantsi:
| Icandelo | Umthamo |
| I-RNA | X µg |
| 10 × DNase I Buffer | 1μL |
| I-DNase I, RNase-free(5U/μL) | I-1 U nge µg RNA① |
| ddH2O | Ukuya kuthi ga kwi-10μL |
Qaphela: ①Bala umthamo we-DNase I ekufuneka ifakwe ngokusekelwe kwisixa se-RNA.
2. 37 ℃ imizuzu eyi-15;
3. Yongeza i-0.5M EDTA kugxininiso lokugqibela lwe-2.5mM ~ 5mM, kunye nokufudumala kwi-65℃ imizuzu eyi-10 ukumisa ukuphendula.Isampulu ingasetyenziswa ngokuthe ngqo kwimpendulo elandelayo njengokukhutshelwa umvaumfuniselo.
Inkcazelo yeyunithi
Iyunithi enye ichazwa njengobungakanani be-enzayimu eya kuthoba ngokupheleleyo i-1µg ye-pBR322I-DNA kwimizuzu eyi-10 kwi-37℃.
Ulawulo lwemeko
RNase:I-5U ye-DNase I ene-1.6µg MS2 RNA kwiiyure ze-4 kwi-37 ℃ ayivelisi ukuthotywa njengokoigqitywe yi-agarose gel electrophoresis.
Amanqaku
1. Nceda ulungiselele i-0.5MEDTA uwedwa.
2. Sebenzisa i-1U DNase I ngeµg ye-RNA.Nangona kunjalo, ukuba i-RNA ingaphantsi kwe-1µg, nceda usebenzise i-1U DNase I.
3. Nceda ubeke i-enzyme emkhenkceni ngexesha lokusebenza.
