DNA I

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.Iṣe ti DNase I da lori Ca2+ ati pe o le muu ṣiṣẹ nipasẹ awọn ions irin divalent gẹgẹbi Mn2+ ati Zn2+.5mM Ca2+ ṣe aabo fun enzymu lati hydrolysis.Ni iwaju Mg2+, enzymu le ṣe idanimọ laileto ati ki o ya aaye eyikeyi lori eyikeyi okun ti DNA.Ni iwaju Mn2+, awọn okun meji ti DNA le jẹ idanimọ nigbakanna ati fifọ ni aaye kanna lati ṣe awọn ajẹkù DNA opin alapin tabi awọn ajẹkù DNA opin alalepo pẹlu awọn nucleotides 1-2 ti n jade.

Ohun-ini Ọja

Bovine Pancreas DNAse Mo jẹ afihan ni eto ikosile iwukara ati mimọ.

Calagbara

Gbigbe ati Ibi ipamọ

1. Iduroṣinṣin Ibi ipamọ: - 15 ℃ ~ -25 ℃ fun ibi ipamọ;

2.Transport Stability: Gbigbe labẹ awọn akopọ yinyin;

3. Ti a pese ni: 10 mM Tris-HCl, 2 mM CaCl₂, 50% glycerol, pH 7.6 ni 25 ℃.

Unit Definition

Ẹyọ kan jẹ asọye bi iye henensiamu eyiti yoo sọ di 1 μg ti pBR322 DNA patapata ni iṣẹju mẹwa 10 ni 37°C.

Iṣakoso didara

RNase:5U ti DNase I pẹlu 1.6 μg MS2 RNA fun wakati 4 ni 37 ℃ ko ni ibajẹ bi a ti pinnu nipasẹ agarose gel electrophoresis.

Kokoro Endotoxin:LAL-igbeyewo, ni ibamu si Chinese Pharmacopoeia IV 2020 àtúnse, jeli iye igbeyewo ọna, gbogboogbo ofin (1143).Akoonu endotoxin kokoro yẹ ki o jẹ ≤10 EU/mg.

Awọn ilana fun Lilo

1.Prepare ojutu idahun ni tube-free RNase ni ibamu si awọn ipin ti a ṣe akojọ si isalẹ:

2.37 ℃ fun iṣẹju 15;

3.Fi ifipamọ ifopinsi lati da iṣesi duro, ati ooru ni 65 ℃ fun awọn iṣẹju 10 lati mu DNase ṣiṣẹ I. Ayẹwo le ṣee lo taara fun idanwo transcription atẹle.

Awọn akọsilẹ

1.Lo 1U DNAse I fun μg ti RNA, tabi 1U DNAse I fun kere ju 1μg ti RNA.

2.EDTA yẹ ki o wa ni afikun si ifọkansi ipari ti 5 mM lati daabobo RNA lati ibajẹ lakoko aiṣiṣẹ enzymu.