DNase I
DNase I (Deoxyribonuclease I) yog ib qho endodeoxyribonuclease uas tuaj yeem zom cov DNA ib leeg lossis ob npaug.Nws paub thiab cleaves phosphodiester bonds los tsim monodeoxynucleotides los yog ib leeg-los yog ob-stranded oligodeoxynucleotides nrog phosphate pawg ntawm 5′-terminal thiab hydroxyl ntawm 3′-terminal.Qhov kev ua ntawm DNase kuv nyob ntawm Ca2+ thiab tuaj yeem qhib tau los ntawm cov hlau sib txawv xws li Mn2+ thiab Zn2+.5mM Ca2+ tiv thaiv cov enzyme los ntawm hydrolysis.Nyob rau hauv lub xub ntiag ntawm Mg2 +, lub enzyme yuav randomly paub thiab cleave txhua qhov chaw ntawm ib tug strand ntawm DNA.Nyob rau hauv lub xub ntiag ntawm Mn2+, ob txoj hlua ntawm DNA tuaj yeem raug lees paub ib txhij thiab cleaved ntawm yuav luag tib lub xaib kom tsim cov kab kawg DNA tawg lossis nplaum kawg DNA tawg nrog 1-2 nucleotides protruding.
Khoom Khoom
Bovine Pancreas DNase Kuv tau hais tawm hauv cov kab mob poov xab thiab ua kom huv.
Ccov omponents
| Cheebtsam | Ntim | |||
| 0.1 TSI | 1 KU | 5 KUB | 50 KUB | |
| DNase I, RNase-dawb | 20 l m | 200 mL | 1 mL ib | 10 ml |
| 10 × DNase Kuv Buffer | 1 mL ib | 1 mL ib | 5 x 1ml | 5 × 10 ml |
Kev thauj mus los thiab khaws cia
1. Cia ruaj khov: -15 ℃ ~ -25 ℃ rau kev cia;
2.Txoj kev ruaj ntseg: Thauj hauv qab ntim dej khov;
3. Muab nyob rau hauv: 10 mM Tris-HCl, 2 mM CaCl250% glycerol, pH 7.6 ntawm 25 ℃.
Unit txhais
Ib chav tsev yog txhais raws li tus nqi ntawm cov enzyme uas yuav ua kom tiav 1 µg ntawm pBR322 DNA hauv 10 feeb ntawm 37 ° C.
Kev Tswj Xyuas Zoo
RNase:5U ntawm DNase I nrog 1.6 μg MS2 RNA rau 4 teev ntawm 37 ℃ yields tsis degradation raws li txiav txim los ntawm agarose gel electrophoresis.
Kab mob Endotoxin:LAL-test, raws li Suav Pharmacopoeia IV 2020 ib tsab, gel limit test method, general rule (1143).Cov kab mob endotoxin cov ntsiab lus yuav tsum yog ≤10 EU / mg.
Cov lus qhia rau kev siv
1. Npaj cov tshuaj tiv thaiv nyob rau hauv RNase-free raj raws li cov proportions teev hauv qab no:
| Cheebtsam | Ntim |
| RNA | X m g |
| 10 × DNase kuv Buffer | 1 mL ib |
| DNase I, RNase-dawb (5U / μL) | 1 U rau μg RNA |
| ddH ua2O | Txog li 10 μL |
2.37 ℃ rau 15 feeb;
3.Ntxiv qhov kev txiav tawm tsis tu ncua kom tsis txhob muaj cov tshuaj tiv thaiv, thiab cua sov ntawm 65 ℃ rau 10 feeb kom tsis txhob siv DNase I. Cov qauv tuaj yeem siv ncaj qha rau kev sim ntsuas tom ntej.
Sau ntawv
1.Siv 1U DNase I ib μg ntawm RNA, lossis 1U DNase I rau tsawg dua 1μg ntawm RNA.
2.EDTA yuav tsum tau ntxiv rau qhov kawg ntawm 5 mM los tiv thaiv RNA los ntawm kev degraded thaum lub sij hawm enzyme inactivation.
