DNase I

ʻO DNase I (Deoxyribonuclease I) he endodeoxyribonuclease hiki ke ʻeli i ka DNA kaula hoʻokahi a i ʻole.Hoʻomaopopo a hoʻokaʻawale i nā 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.Aia ka hana o DNase I ma Ca2+ a hiki ke ho'āla ʻia e nā ion metala divalent e like me Mn2+ a me Zn2+.Mālama ʻo 5mM Ca2+ 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 ʻelua 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 pili me 1-2 nucleotides e puka mai ana.

Waiwai Huahana

ʻO Bovine Pancreas DNase I hōʻike ʻia i ka ʻōnaehana hōʻike hū a hoʻomaʻemaʻe ʻia.

Cnā mea hoʻokūkū

ʻO ka lawe ʻana a me ka mālama ʻana

1. Paʻa Paʻa: - 15 ℃ ~ -25 ℃ no ka mālama ʻana;

2.Transport Stability: ʻO ka halihali ma lalo o nā pahu hau;

3. Hāʻawi ʻia ma: 10 mM Tris-HCl, 2 mM CaCl₂, 50% glycerol, pH 7.6 ma 25 ℃ .

Wehewehe Unite

Hoʻokahi ʻāpana i wehewehe ʻia ʻo ka nui o ka enzyme e hoʻohaʻahaʻa loa i 1 µg o pBR322 DNA i 10 mau minuke ma 37°C.

Ka Mana Mana

RNase:ʻO 5U o DNase I me 1.6 μg MS2 RNA no 4 mau hola ma 37 ℃ ʻaʻole e hāʻawi i ka degradation e like me ka hoʻoholo ʻana e ka electrophoresis gel agarose.

Bakteri Endotoxin:LAL-hōʻike, e like me ka Chinese Pharmacopoeia IV 2020 edition, gel limit test method, general rule (1143).Pono ka maʻiʻo endotoxin bacteria ≤10 EU/mg.

Nā kuhikuhi no ka hoʻohana ʻana

1. E hoʻomākaukau i ka hopena hopena i loko o ka pahu RNase-free e like me nā ʻāpana i helu ʻia ma lalo nei.

2.37 ℃ no 15 mau minuke;

3. E hoʻohui i ka pahu hoʻopau e hoʻopau i ka hopena, a me ka wela ma 65 ℃ no nā minuke 10 e hoʻopau i ka DNase I. Hiki ke hoʻohana pololei ʻia ka hāpana no ka hoʻokolohua transcription aʻe.

Nā memo

1. E hoʻohana i ka 1U DNase I no μg o RNA, a i ʻole 1U DNase I no ka liʻiliʻi o 1μg o RNA.

Pono e hoʻohui ʻia ʻo 2.EDTA i ka hopena hope o 5 mM e pale i ka RNA mai ka hoʻohaʻahaʻa ʻana i ka wā o ka enzyme inactivation.