DNAse I (Rnase Ọfẹ)(5u/ul)
Nọmba ologbo: HC4007A
DNase I (Deoxyribonuclease I) jẹ ẹya endodeoxyribonuclease ti o le di ẹyọkan tabi DNA ti o ni okun meji.O mọ ati cleaves phosphodiester bonds lati gbe awọn monodeoxynucleotides tabi nikan- tabi ilopo-strand oligodeoxynucleotides pẹlu fosifeti awọn ẹgbẹ ni 5'-terminal ati hydroxyl ni 3'-terminal.Iṣẹ-ṣiṣe ti DNAse I da lori Ca2+ati pe o le muu ṣiṣẹ nipasẹ awọn ions irin divalent gẹgẹbi Mn2+ati Zn2+.5 mM Ca2+ṣe aabo fun enzymu lati hydrolysis.Ni iwaju ti Mg2+, enzymu le ṣe idanimọ laileto ati ki o di aaye eyikeyi lori eyikeyi okun ti DNA.Niwaju Mn2+, awọn okun meji ti DNA le jẹ idanimọ nigbakanna ati fifọ ni fere aaye kanna lati ṣe awọn ajẹkù DNA opin alapin tabi awọn ajẹkù DNA opin alalepo pẹlu awọn nucleotides 1-2 ti n jade.
Awọn eroja
| Oruko | 0.1KU | 1KU | 5 KU | 50 KU |
| DNAse I, RNase-ọfẹ | 20μL | 200μL | 1ml | 10 milimita |
| 10×DNase Mo saarin | 1ml | 1ml | 5 × 1 milimita | 5 ×10 milimita |
Awọn ipo ipamọ
-25 ℃ ~ -15 ℃ fun ibi ipamọ;Transport labẹ yinyin akopọ.
Awọn ilana
1. Mura ojutu ifaseyin ninu tube ti ko ni RNase ni ibamu si awọn iwọn ti a ṣe akojọ si isalẹ:
| Ẹya ara ẹrọ | Iwọn didun |
| RNA | X µg |
| 10 × DNAse Mo Ifipamọ | 1μL |
| DNAase I, RNase-ọfẹ (5U/μL) | 1 U fun µg RNA① |
| ddH2O | Titi di 10 μL |
Akiyesi: ① Ṣe iṣiro iwọn didun DNAse I ti o nilo lati ṣafikun da lori iye RNA.
2. 37 ℃ fun iṣẹju 15;
3. Fi 0.5M EDTA kun si ifọkansi ikẹhin ti 2.5mM ~ 5mM, ati ooru ni 65 ℃ fun awọn iṣẹju 10 lati da iṣesi naa duro.Apeere naa le ṣee lo taara fun iṣesi atẹle gẹgẹbi iṣipopada transcriptionṣàdánwò.
Unit Definition
Ẹyọ kan jẹ asọye bi iye henensiamu eyiti yoo sọ di 1µg ti pBR322 patapataDNA ni iṣẹju mẹwa 10 ni 37 ℃.
Iṣakoso didara
RNase:5U ti DNAse I pẹlu 1.6µg MS2 RNA fun awọn wakati 4 ni 37℃ ko ni ibajẹ bipinnu nipasẹ agarose gel electrophoresis.
Awọn akọsilẹ
1. Jọwọ mura 0.5MEDTA nipasẹ ara rẹ.
2. Lo 1U DNAse I fun µg ti RNA.Sibẹsibẹ, ti RNA ba kere ju 1µg, jọwọ lo 1U DNAse I.
3. Jọwọ gbe enzymu sori yinyin nigba iṣẹ.
