DNase I (Rnase Koreutu)(5u/ul)
Cat Nama: HC4007A
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 hononga phosphodiester ki te whakaputa i te monodeoxynucleotides, te oligodeoxynucleotides kotahi, takirua ranei me nga roopu phosphate i te 5'-terminal me te hydroxyl i te 3'-terminal.Ko te mahi a DNase I ka whakawhirinaki ki te Ca2+a ka taea te whakahohe e nga katote whakarewa divalent penei i a Mn2+me Zn2+.5 mM Ca2+he whakamarumaru i te enzyme mai i te hauwai.I te aroaro o 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+, Ko nga whenu rua o te DNA ka taea te mohio i te wa kotahi ka wehea ki te waahi kotahi hei hanga i nga kongakonga DNA mutunga papatahi, nga kongakonga DNA mutunga piri me te 1-2 nucleotides e puta ana.
Waehanga
| Ingoa | 0.1KU | 1KU | 5 KU | 50 KU |
| DNAase I, RNase-kore | 20μL | 200μL | 1mL | 10 mL |
| 10×DNase ahau Kaipupuri | 1mL | 1mL | 5 × 1mL | 5 ×10 mL |
Nga tikanga rokiroki
-25 ℃ ~ -15 ℃ mo te rokiroki;Te kawe i raro i nga kete hukapapa.
Tohutohu
1. Whakapaia te otinga tauhohenga i roto i te ngongo kore-Rnase kia rite ki nga waahanga kua whakarārangitia i raro nei:
| Waehanga | Rōrahi |
| RNA | X µg |
| 10 × DNase I Kaipupuri | 1μL |
| DNase I, RNase-kore(5U/μL) | 1 U mo ia µg RNA① |
| ddH2O | Tae atu ki te 10μL |
Kia mahara: ①Tātaihia te rōrahi o te DNase I me tāpiri i runga i te nui o te RNA.
2. 37 ℃ mō te 15 meneti;
3. Tāpirihia te 0.5M EDTA ki te kukū whakamutunga o te 2.5mM~5mM, ka whakamahana i te 65 ℃ mo te 10 meneti hei whakamutu i te tauhohenga.Ka taea te whakamahi tika i te tauira mo te tauhohenga e whai ake nei penei i te tuhi whakamuriwhakamātau.
Whakamaramatanga Waeine
Kotahi te waeine e kiia ana ko te nui o te whaakiki ka tino whakaheke i te 1µg o pBR322DNA i roto i te 10 meneti i te 37 ℃.
Mana Kounga
RNase:Ko te 5U o DNase I me te 1.6µg MS2 RNA mo te 4 haora i te 37 ℃ karekau he whakahekeka whakatauhia e te electrophoresis gel agarose.
Notes
1. Tena koa whakarite 0.5MEDTA koe.
2. Whakamahia te 1U DNase I mo ia µg o te RNA.Heoi, ki te iti iho te RNA i te 1µg, whakamahia te 1U DNase I.
3. Tuhia koa te enzyme ki runga tio i te wa e mahi ana.
