DNase I
DNase I (Deoxyribonuclease I) ke endodeoxyribonuclease e ka silang DNA e nang le mela e le 'ngoe kapa e habeli.E lemoha le ho qhetsola litlamo tsa phosphodiester ho hlahisa monodeoxynucleotides kapa oligodeoxynucleotide e nang le stranded e le 'ngoe kapa habeli ka lihlopha tsa phosphate sebakeng sa 5'-terminal le hydroxyl ho 3'-terminal.Ts'ebetso ea DNase I e ipapisitse le Ca2 + mme e ka ts'oaroa ke li-ion tsa tšepe tsa divalent joalo ka Mn2+ le Zn2+.5mM Ca2+ e sireletsa enzyme ho tsoa ho hydrolysis.Ha Mg2+ e le teng, enzyme e ne e khona ho lemoha le ho petsola sebaka leha e le sefe sa DNA.Ka pel'a Mn2+, likhoele tse peli tsa DNA li ka hlokomeloa ka nako e le 'ngoe' me tsa phunyeletsoa sebakeng se le seng ho etsa likaroloana tsa DNA kapa likhechana tsa DNA tse nang le 1-2 nucleotide.
Thepa ea Sehlahisoa
Bovine Pancreas DNase I e ile ea hlahisoa ka mokhoa oa polelo ea tomoso mme ea hloekisoa.
Cbahanyetsi
| Karolo | Bolumo | |||
| 0.1KU | 1KU | 5KU | 50KU | |
| DNase I, RNase-mahala | 20μL | 200μL | 1mL | 10mL |
| 10×DNase I Buffer | 1mL | 1mL | 5 × 1mL | 5 × 10mL |
Lipalangoang le Polokelo
1. Storage Stability: – 15℃~-25℃ bakeng sa polokelo;
2.Botsitso ba Lipalangoang: Lipalangoang tlas'a mekotla ea leqhoa;
3. E fanoe ka: 10 mM Tris-HCl, 2 mM CaCl2, 50% glycerol, pH 7.6 ho 25℃.
Tlhaloso ea Yuniti
Yuniti e le 'ngoe e hlalosoa e le palo ea enzyme e tla theola 1 µg ea pBR322 DNA ka ho felletseng ka metsotso e 10 ho 37°C.
Taolo ea Boleng
RNase:5U ea DNase I e nang le 1.6 μg MS2 RNA bakeng sa lihora tse 4 ho 37 ℃ ha e fane ka ho senyeha ho ea ka qeto ea agarose gel electrophoresis.
Baktheria Endotoxin:Tlhahlobo ea LAL, ho latela khatiso ea China Pharmacopoeia IV 2020, mokhoa oa tlhahlobo ea moeli oa gel, molao o akaretsang (1143).Likahare tsa endotoxin ea baktheria e lokela ho ba ≤10 EU/mg.
Litaelo tsa ho Sebelisa
1. Lokisetsa tharollo ea karabelo ka har'a tube e se nang RNase ho latela likarolo tse thathamisitsoeng ka tlase:
| Karolo | Bolumo |
| RNA | X μg |
| 10 × DNase I Buffer | 1 μL |
| DNase I, RNase-mahala(5U/μL) | 1 U ka μg RNA |
| ddH2O | Ho fihlela ho 10 μL |
2.37 ℃ metsotso e 15;
3.Eketsa buffer ea ho emisa ho emisa karabelo, 'me u chese ka 65℃ metsotso e 10 ho tima DNase I. Sampole e ka sebelisoa ka kotloloho bakeng sa teko e latelang ea transcript.
Lintlha
1.Sebelisa 1U DNase I ka μg ea RNA, kapa 1U DNase I ka tlase ho 1μg ea RNA.
2.EDTA e lokela ho kenngoa sebakeng sa ho qetela sa 5 mM ho sireletsa RNA hore e se ke ea senyeha nakong ea ho se sebetse ha enzyme.
