DNase I
Ko te DNase I (Deoxyribonuclease I) he endodeoxyribonuclease ka taea te keri i te DNA whenu kotahi-rua ranei.Ka mohio, ka wehe i nga here phosphodiester ki te whakaputa i te monodeoxynucleotides, oligodeoxynucleotides kotahi-rua ranei me nga roopu phosphate i te 5'-terminal me te hydroxyl i te 3'-terminal.Ko te ngohe o DNase I kei runga i te Ca2+ ka taea te whakahohe e nga katote whakarewa divalent penei Mn2+ me Zn2+.Ko te 5mM Ca2+ he whakamarumaru i te whākōkī mai i te hauwai.I te aroaro o te Mg2+, ka taea e te enzyme te mohio matapōkeretia me te wawahi i tetahi waahi i runga i tetahi whenu DNA.I te aroaro o Mn2+, ka taea te mohio nga whenu rua o te DNA i te wa kotahi, ka wehea ki te waahi kotahi hei hanga i nga kongakonga DNA mutunga papatahi, i nga kongakonga DNA mutunga piri me te 1-2 nucleotides e puta ana.
Taonga Hua
Bovine Pancreas DNase I whakaatuhia ahau i roto i te punaha whakaputa rewena me te purea.
Components
| Waehanga | Rōrahi | |||
| 0.1KU | 1KU | 5KU | 50KU | |
| DNAase I, RNase-kore | 20μL | 200μL | 1mL | 10mL |
| 10×DNase ahau Kaipupuri | 1mL | 1mL | 5×1mL | 5×10mL |
Te kawe me te rokiroki
1. Rokiroki Rokiroki: - 15 ℃ ~ -25 ℃ mo te rokiroki;
2.Transport Stability: Te kawe i raro i nga peke tio;
3. Ka tukuna ki roto: 10 mM Tris-HCl, 2 mM CaCl2, 50% glycerol, pH 7.6 i te 25 ℃.
Whakamaramatanga Waeine
Kotahi te waeine kua tautuhia ko te nui o te whaakiki ka tino whakaheke i te 1 µg o pBR322 DNA i roto i te 10 meneti i te 37°C.
Mana Kounga
RNase:Ko te 5U o te DNase I me te 1.6 μg MS2 RNA mo te 4 haora i te 37 ℃ karekau he paheketanga ka whakatauhia e te electrophoresis gel agarose.
huakita Endotoxin:LAL-whakamatautau, e ai ki te Hainamana Pharmacopoeia IV 2020 whakaputanga, tikanga whakamatautau tepe gel, ture whanui (1143).Ko te ihirangi endotoxin huakita kia ≤10 EU/mg.
Tohutohu mo te Whakamahi
1.Whakaritea te otinga tauhohenga i roto i te ngongo kore utu RNase kia rite ki nga waahanga kua whakarārangitia i raro nei:
| Waehanga | Rōrahi |
| RNA | X μg |
| 10 × DNase I Kaipupuri | 1 μL |
| DNase I, RNase-kore(5U/μL) | 1 U mo ia μg RNA |
| ddH2O | Tae atu ki te 10 μL |
2.37 ℃ mō te 15 meneti;
3. Tāpirihia te parepare whakamutu ki te whakamutu i te tauhohenga, me te wera i te 65 ℃ mo te 10 meneti hei whakakore i te DNase I. Ka taea te whakamahi tika i te tauira mo te whakamatautau tuhinga e whai ake nei.
Notes
1.Whakamahia 1U DNase I ia μg o RNA, 1U DNase I ranei mo te iti iho i te 1μg o RNA.
2.Me taapiri te EDTA ki te kukū whakamutunga o te 5 mM hei tiaki i te RNA kia kore e paheke i te wa e kore ai e whakahohehia te enzyme.
