DNase I (Rnase Yemahara) (2u/ul)
Katsi Nhamba: HC4007B
DNase I is endonuclease inokwanisa kugaya DNA imwe-kana kaviri-tambo.Inogona hydrolyze phosphodiester bonds kugadzira mono-uye oligodeoxynucleotides ine 5'-phosphate boka uye 3'-OH boka.Iyo yakakwana yekushanda pH huwandu hweDNase I ndeye 7-8.Chiitiko cheDNase I chinoenderana neCa2+uye inogona kuitwa ne divalent simbi ions senge CO2, Mn2+, Zn2+, etc. Pamberi paMg2+, DNase Ndinogona kutsemura chero nzvimbo yeDNA ine tambo mbiri;Ndichiri pamberi paMn2+, DNase Ndinokwanisa kutsemura DNA yakapetwa kaviri panzvimbo imwe chete, ichiita migumo yakagomara kana inonamira migumo ine 1-2 nucleotides yakabudikira.Inogona kushandiswa kugadzirisa akasiyana siyana eRNA samples.
Zvikamu
| Zita | 1KU | 5KU |
| Recombinant DNaseI (RNase-isina) | 500 μL | 5 ×500 μL |
| DNase I Reaction Buffer (10×) | 1 mL | 5 × 1mL |
Kuchengetedza mamiriro
Ichi chigadzirwa chinofanira kuchengetwa pa -25 ~ -15 ℃ kwemakore maviri.Ndapota dzivisa kuomesa nechando kakawanda.
Mirayiridzo
Inoshandiswa pakubviswa kweDNA kubva kuRNA samples yekutarisa chete.
1. Ndokumbira ushandise RNase-isina centrifuge machubhu nemapipette matipi kugadzirira inotevera maitiro system:
| Zvikamu | Vhoriyamu (μL) |
| DNase I Reaction Buffer (10×) | 1 |
| Recombinant DNasel (RNase-isina) | 1 |
| RNA | X |
| RNase-isina ddH2O | Kusvikira ku10 |
2. Mamiriro ekugadzirisa maitiro ndeaya anotevera: 37 ℃, mushure me15-30 mins, wedzera mutsara wekupedzisira we 2.5 mM EDTA mhinduro uye funganisa zvakanaka, ipapo 65 ℃ kwemaminitsi gumi.Iyo template yakagadziriswa inogona kushandiswa kune inotevera RT-PCR kana RT-qPCR kuyedza, nezvimwe.
Notes
1. DNase l inonzwa kune denaturation yemuviri;Paunenge uchivhenganisa, zvinyoronyoro dzosera test tube uyezunza zvakanaka, usazunungusa nesimba.
2. Iyo enzyme inofanira kuchengetwa mubhokisi rechando kana pamvura yechando kana yakashandiswa, uye inofanira kuchengetwa pa -20 ℃ pakarepo mushure mekushandisa.
3. Ichi chigadzirwa ndechekutsvaga kushandiswa chete.
4. Ndokumbira ushande nemajasi erabhu uye magirovhosi anoraswa, kuti uchengetedzeke.
