DNA I
DNase I (Deoxyribonuclease I) wani endodeoxyribonuclease ne wanda zai iya narkar da DNA guda ɗaya ko biyu.Yana gane kuma yana raba haɗin phosphodiester don samar da monodeoxynucleotides ko oligodeoxynucleotides guda ɗaya ko biyu tare da ƙungiyoyin phosphate a 5'-terminal da hydroxyl a 3'-terminal.Ayyukan DNAse I ya dogara da Ca2+ kuma ana iya kunna su ta hanyar ions na ƙarfe daban-daban kamar Mn2+ da Zn2+.5mM Ca2+ yana kare enzyme daga hydrolysis.A gaban Mg2+, enzyme na iya ganewa da kayyade kuma ya raba kowane rukunin yanar gizo akan kowane madaidaicin DNA.A gaban Mn2+, za'a iya gane nau'ikan DNA guda biyu lokaci guda kuma a raba su a kusan wuri ɗaya don samar da gutsuttsuran ƙarshen DNA ko gutsuttsuran ƙarshen DNA tare da 1-2 nucleotides suna fitowa.
Kayayyakin Samfura
Bovine Pancreas DNase An bayyana ni a tsarin bayyana yisti kuma an tsarkake ni.
Cmasu kai hari
| Bangaren | Ƙarar | |||
| 0.1 KU | 1 KU | 5KU | 50 KU | |
| DNSase I, RNase-free | 20 μl | 200 μL | 1 ml | ml 10 |
| 10×DNase I Buffer | 1 ml | 1 ml | 5 × 1 ml | 5 × 10 ml |
Sufuri da Ajiya
1. Storage Stability: - 15 ℃ ~ -25 ℃ don ajiya;
2.Transport Stability: Transport a karkashin fakitin kankara;
3. An ba da shi: 10 mM Tris-HCl, 2 mM CaCl250% glycerol, pH 7.6 a 25 ℃.
Ma'anar Naúrar
An bayyana raka'a ɗaya azaman adadin enzyme wanda zai lalata 1 μg na pBR322 DNA gaba ɗaya cikin mintuna 10 a 37°C.
Kula da inganci
RNase:5U na DNase I tare da 1.6 μg MS2 RNA na tsawon awanni 4 a 37 ℃ ba ya haifar da lalacewa kamar yadda aka ƙaddara ta agarose gel electrophoresis.
Kwayoyin cuta Endotoxin:LAL-gwajin, bisa ga Pharmacopoeia IV 2020 na Sinanci, hanyar gwajin iyaka ta gel, ƙa'ida ta gaba ɗaya (1143).Ya kamata abun cikin endotoxin na ƙwayoyin cuta ya zama ≤10 EU/mg.
Umarnin don Amfani
1.Shirya maganin amsawa a cikin bututu mai kyauta na RNase bisa ga adadin da aka jera a ƙasa:
| Bangaren | Ƙarar |
| RNA | X μg |
| 10 × DNSase I Buffer | 1 μl |
| DNSase I, RNase-kyauta(5U/μL) | 1 U da μg RNA |
| ddH2O | Har zuwa 10 μl |
2.37 ℃ na minti 15;
3.Ƙara buffer ƙarewa don dakatar da amsawa, kuma zafi a 65 ℃ na minti 10 don kunna DNAse I. Za'a iya amfani da samfurin kai tsaye don gwaji na gaba na gaba.
Bayanan kula
1.Yi amfani da 1U DNAse I a kowace μg na RNA, ko 1U DNAse I don ƙasa da 1μg na RNA.
2.EDTA ya kamata a ƙara zuwa ƙaddamarwa na ƙarshe na 5 mM don kare RNA daga lalacewa yayin rashin kunna enzyme.
