DNase I (Rnase Ubuntu) (5u / ul)
Injangwe No: HC4007A
DNase I (Deoxyribonuclease I) ni endodeoxyribonuclease ishobora gusya ADN imwe cyangwa ebyiri.Iremera kandi ikomatanya imiyoboro ya fosifori kugirango itange monodeoxynucleotide cyangwa oligodeoxynucleotide imwe cyangwa ebyiri-ebyiri hamwe na fosifate mumatsinda ya 5'-terminal na hydroxyl kuri 3'-terminal.Igikorwa cya DNase I giterwa na Ca.2+kandi irashobora gukoreshwa na ion zingana nicyuma nka Mn2+na Zn2+.5 mM Ca.2+irinda enzyme hydrolysis.Imbere ya Mg2+, enzyme yashoboraga kumenya kubushake no gutobora urubuga urwo arirwo rwose rwa ADN.Imbere ya Mn2+, imirongo ibiri ya ADN irashobora kumenyekana icyarimwe kandi igashyirwa kumurongo hafi imwe kugirango ibe ibice bya ADN bigororotse cyangwa ibice bya ADN bifatanye hamwe na nucleotide 1-2.
Ibigize
| Izina | 0.1KU | 1KU | 5 KU | 50 KU |
| DNase I, RNase-yubusa | 20μL | 200μL | 1mL | 10 mL |
| 10 × DNase I Buffer | 1mL | 1mL | 5 × 1mL | 5 × 10 mL |
Imiterere yo kubika
-25 ℃ ~ -15 ℃ kubika;Ubwikorezi munsi yububiko.
Amabwiriza
1. Tegura igisubizo cya reaction muri tube idafite RNase ukurikije ibipimo byavuzwe hepfo:
| Ibigize | Umubumbe |
| RNA | X µg |
| 10 × DNase I Buffer | 1μL |
| DNase I, RNase-yubusa (5U / μL) | 1 U kuri µg RNA① |
| ddH2O | Kugera kuri 10μL |
Icyitonderwa: alBara ingano ya DNase I igomba kongerwaho ukurikije ingano ya RNA.
2. 37 ℃ mu minota 15;
3. Ongeramo 0.5M EDTA kumurongo wanyuma wa 2.5mM ~ 5mM, hanyuma ushushe kuri 65 ℃ muminota 10 kugirango uhagarike reaction.Icyitegererezo kirashobora gukoreshwa muburyo butaziguye nka reaction ikurikiraigerageza.
Igisobanuro
Igice kimwe gisobanurwa nkubunini bwa enzyme izatesha agaciro 1µg ya pBR322ADN mu minota 10 kuri 37 ℃.
Kugenzura ubuziranenge
RNase:5U ya DNase I hamwe na 1.6µg MS2 RNA kumasaha 4 kuri 37 ℃ nta musaruro utanga nkukobigenwa na agarose gel electrophorei.
Inyandiko
1. Nyamuneka tegura 0.5MEDTA wenyine.
2. Koresha 1U DNase I kuri µg ya RNA.Ariko, niba RNA iri munsi ya 1µg, nyamuneka koresha 1U DNase I.
3. Nyamuneka shyira enzyme kurubura mugihe ukora.
