DNase I (Rnase Free)(5u/ul)
Helu popoki: HC4007A
ʻO DNase I (Deoxyribonuclease I) he endodeoxyribonuclease hiki ke ʻeli i ka DNA kaula hoʻokahi a i ʻole.Hoʻomaopopo a hoʻokaʻawale i nā mea paʻa phosphodiester e hana i nā monodeoxynucleotides a i ʻole nā oligodeoxynucleotides kaulua a i ʻole me nā hui phosphate ma ka 5'-terminal a me ka hydroxyl ma ka 3'-terminal.ʻO ka hana o DNase I pili iā Ca2+a hiki ke ho'ā 'ia e divalent metala ion e like me Mn2+a me Zn2+.5 mM Ca2+pale i ka enzyme mai ka hydrolysis.Ma ke alo o Mg2+, hiki i ka enzyme ke hoʻomaopopo a hoʻokaʻawale i kekahi kahua ma kekahi kaula DNA.Ma ke alo o Mn2+, Hiki ke ʻike ʻia nā kaula pālua o ka DNA i ka manawa like a ʻoki ʻia ma kahi kokoke i ka pae hoʻokahi e hana i nā ʻāpana DNA palahalaha a i ʻole nā ʻāpana DNA hope paʻa me 1-2 nucleotides e puka mai ana.
Nā ʻāpana
| inoa | 0.1KU | 1KU | 5 KU | 50 KU |
| DNase I, RNase-noa | 20μL | 200μL | 1mL | 10 mL |
| 10×DNase I Buffer | 1mL | 1mL | 5 × 1mL | 5 ×10 mL |
Nā kūlana mālama
-25 ℃ ~ -15 ℃ no ka mālama ʻana;Lawe ʻia ma lalo o nā ʻeke hau.
Nā kuhikuhi
1. E hoʻomākaukau i ka hoʻonā hopena i loko o ka paipu RNase-free e like me nā ʻāpana i helu ʻia ma lalo nei.
| ʻāpana | Volume |
| RNA | X µg |
| 10 × DNase I Buffer | 1μL |
| DNase I, RNase-noa (5U/μL) | 1 U no µg RNA① |
| ddH2O | A hiki i 10μL |
'Ōlelo Aʻo: ①E helu i ka nui o DNase I pono e hoʻohui ʻia ma muli o ka nui o RNA.
2. 37 ℃ no 15 mau minuke;
3. E hoʻohui i ka 0.5M EDTA i ka hopena hope loa o 2.5mM ~ 5mM, a me ka wela ma 65 ℃ no 10 mau minuke e hoʻopau i ka hopena.Hiki ke hoʻohana pololei ʻia ka hāpana no ka hopena aʻe e like me ka hoʻohuli ʻanahoʻokolohua.
Wehewehe Unite
Hoʻokahi ʻāpana i wehewehe ʻia ʻo ka nui o ka enzyme e hoʻohaʻahaʻa loa i 1µg o pBR322DNA i 10 mau minuke ma 37 ℃.
Ka Mana Mana
RNase:ʻO 5U o DNase I me 1.6µg MS2 RNA no 4 mau hola ma 37 ℃ ʻaʻole e hāʻawi i ka hoʻohaʻahaʻa.Hoʻoholo ʻia e ka electrophoresis gel agarose.
Nā memo
1. E ʻoluʻolu e hoʻomākaukau iā 0.5MEDTA iā ʻoe iho.
2. E hoohana i 1U DNase I no µg o RNA.Eia naʻe, inā emi ka RNA ma mua o 1µg, e ʻoluʻolu e hoʻohana i ka 1U DNase I.
3. E ʻoluʻolu e kau i ka enzyme ma luna o ka hau i ka wā e hana ai.
