DNase I (Rnase Yemahara)(5u/ul)
Katsi Nhamba: HC4007A
DNase I (Deoxyribonuclease I) inonzi endodeoxyribonuclease inokwanisa kugaya DNA imwe kana ine tambo mbiri.Inoziva uye inotsemura mabhondi ephosphodiester kugadzira monodeoxynucleotides kana oligodeoxynucleotides imwe kana kaviri-yakasungwa nemapoka ephosphate pa5'-terminal uye hydroxyl pa3'-terminal.Chiitiko cheDNase I chinoenderana neCa2+uye inogona kuitwa ne divalent simbi ions senge Mn2+uye zn2+.5 mM Ca2+inodzivirira iyo enzyme kubva kune hydrolysis.Pamberi paMg2+, enzyme yaigona kungoziva uye kutsemura chero nzvimbo pane chero tambo yeDNA.Pamberi paMn2+, tambo mbiri dzeDNA dzinogona kuzivikanwa panguva imwe chete uye dzinotsemurwa panzvimbo imwe chete kuita zvidimbu zveDNA zvakati sandarara kana kuti zvimedu zveDNA zvinonamira zvine 1-2 nucleotides.
Zvikamu
| Zita | 0.1KU | 1KU | 5 KU | 50 KU |
| DNase I, RNase-isina | 20μL | 200μL | 1mL | 10 mL |
| 10 × DNase I Buffer | 1mL | 1mL | 5 × 1mL | 5 × 10 mL |
Kuchengetedza mamiriro
-25 ℃~-15 ℃ yekuchengetedza;Kutakura pasi pemazaya echando.
Mirayiridzo
1. Gadzirira mhinduro muRNase-isina chubhu zvinoenderana nechiyero chakanyorwa pazasi:
| Chikamu | Volume |
| RNA | X µg |
| 10 × DNase I Buffer | 1μL |
| DNase I, RNase-isina(5U/μL) | 1 U per µg RNA① |
| ddH2O | Kusvika ku10μL |
Cherechedza: ① Verenga huwandu hweDNase I inoda kuwedzerwa zvichienderana nehuwandu hweRNA.
2. 37 ℃ kwemaminitsi gumi nemashanu;
3. Wedzera 0.5M EDTA kusvika kumagumo ekupedzisira e2.5mM ~ 5mM, uye kupisa pa 65 ℃ kwemaminitsi gumi kuti umise maitiro.Muenzaniso unogona kushandiswa zvakananga kune inotevera kuita senge reverse transcriptionexperiment.
Chikamu Tsanangudzo
Chimwe chikamu chinotsanangurwa sehuwandu hwe enzyme iyo inozosvibisa zvachose 1µg yepBR322DNA mumaminetsi gumi pa37 ℃.
Quality Control
RNase:5U yeDNase I ine 1.6µg MS2 RNA kwemaawa mana pa37 ℃ haiburitseinotsanangurwa neagarose gel electrophoresis.
Notes
1. Ndapota gadzirira 0.5MEDTA iwe pachako.
2. Shandisa 1U DNase I paµg yeRNA.Nekudaro, kana iyo RNA iri pasi pe1µg, ndapota shandisa 1U DNase I.
3. Ndapota isa enzyme pachando panguva yekushanda.
