DNSase I (Rnase Kyauta) (2u/ul)
Saukewa: HC4007B
DNAse I shine endonuclease wanda zai iya narkar da DNA mai ɗaci ɗaya ko biyu.Yana iya hydrolyze phosphodiester bond don samar da mono-da oligodeoxynucleotides dauke da rukunin 5'-phosphate da ƙungiyar 3'-OH.Mafi kyawun kewayon pH mai aiki na DNAse I shine 7-8.Ayyukan DNAse I ya dogara da Ca2+kuma ana iya kunna ta ta hanyar ion ƙarfe na divalent kamar CO2, Mn2+, Zan2+, da sauransu A gaban Mg2+, DNase Zan iya bazuwar kowane rukunin yanar gizo na DNA mai ɗaure biyu;Yayin da yake gaban Mn2+, DNase Zan iya raba DNA guda biyu a wuri guda, samar da ƙullun ƙarewa ko ƙarewa tare da 1-2 nucleotides suna fitowa.Ana iya amfani dashi don sarrafa samfuran RNA daban-daban.
Abubuwan da aka gyara
| Suna | 1 KU | 5KU |
| Recombinant DNSaseI (RNase-free) | 500 μl | 5 × 500 ml |
| DNAase I Reaction Buffer (10×) | 1 ml | 5 × 1 ml |
Yanayin ajiya
Wannan samfurin ya kamata a adana a -25 ~ -15 ℃ for 2 shekaru.Da fatan za a guje wa daskare da aka maimaita.
Umarni
Aiwatar da cire DNA daga samfuran RNA don tunani kawai.
1. Da fatan za a yi amfani da bututun centrifuge marasa kyauta na RNase da tukwici na pipette don shirya tsarin amsa mai zuwa:
| Abubuwan da aka gyara | girma (μL) |
| DNAase I Reaction Buffer (10×) | 1 |
| Recombinant DNasel (RNase-kyauta) | 1 |
| RNA | X |
| ddH mara amfani2O | Har zuwa 10 |
2. A dauki yanayi ne kamar haka: 37 ℃, bayan 15-30 mins, ƙara wani karshe taro na 2.5 mM EDTA bayani da Mix da kyau, sa'an nan 65 ℃ for 10 mins.Ana iya amfani da samfurin da aka sarrafa don gwaje-gwajen RT-PCR ko RT-qPCR na gaba, da sauransu.
Bayanan kula
1. DNase l yana kula da denaturation na jiki;Lokacin haɗuwa, a hankali juya bututun gwajin kumagirgiza da kyau, kar a girgiza da karfi.
2. Dole ne a adana enzymes a cikin akwati na kankara ko kuma a kan wanka na kankara lokacin amfani da shi, kuma ya kamata a adana shi a -20 ℃ nan da nan bayan amfani.
3. Wannan samfurin don amfanin bincike ne kawai.
4. Da fatan za a yi aiki da riguna na lab da safofin hannu na zubarwa, don amincin ku.
