DNAse I (Rnase Bilaash)(5u/ul)
Bisadda No: HC4007A
DNase I (Deoxyribonuclease I) waa endodeoxyribonuclease kaas oo dheefshiidi kara hal-ama laba-geesood DNA.Waxay aqoonsataa oo kala jeexdaa bonds phosphodiester si ay u soo saarto monodeoxynucleotides ama oligodeoxynucleotides hal-ama labajibbaaran oo leh kooxaha fosfatka ee 5'-terminal iyo hydroxyl ee 3'-terminal.Hawlaha DNAse I waxay kuxirantahay Ca2+waxaana lagu hawlgelin karaa ion biraha kala duwan sida Mn2+iyo Zn2+.5 mM Ca2+waxay ka ilaalisaa enzyme ka hydrolysis.Iyadoo uu joogo Mg2+, Insaymku wuxuu si aan kala sooc lahayn u aqoonsan karaa oo u kala goyn karaa goob kasta oo ku taal qayb kasta oo DNA ah.Isagoo goob joog ka ahaa Mn2+, labada qaybood ee DNA-da ayaa isku mar la aqoonsan karaa oo la kala dillaaci karaa ku dhawaad isla goobta si ay u sameeyaan jajabyo DNA-da oo siman ama jajab dhammaadka dhegdhegaysan ee DNA oo leh 1-2 nucleotide.
Qaybaha
| Magaca | 0.1KU | 1KU | 5 KU | 50 KU |
| DNAse I, RNase-free | 20μL | 200μL | 1ml | 10 ml |
| 10×DNase I Buffer | 1ml | 1ml | 5 × 1ml | 5 × 10 ml |
Xaaladaha kaydinta
-25 ℃ ~ -15 ℃ kaydinta;Gaadiidka hoostooda xirmooyinka barafka.
Tilmaamaha
1. U diyaari xalka falcelinta tuubada-free RNase sida waafaqsan saamiga hoos ku taxan:
| Qayb | Mugga |
| RNA | X µg |
| 10 × DNase I Buffer | 1μL |
| DNA-ga I, RNase-free (5U/μL) | 1 U halkii µg RNA① |
| ddH2O | Ilaa 10μL |
Xusuusin: ① Xisaabi mugga DNA-ga I ee u baahan in lagu daro iyadoo lagu salaynayo cadadka RNA.
2. 37 ℃ 15 daqiiqo;
3. Ku dar 0.5M EDTA heerka ugu dambeeya ee 2.5mM~5mM, kuleylna 65℃ 10 daqiiqo si aad u joojiso falcelinta.Muunada waxaa si toos ah loogu isticmaali karaa falcelinta soo socota sida qoraal-ku-noqoshotijaabo.
Qeexitaan Unug
Hal unug ayaa lagu qeexaa cadadka ensaymka kaas oo gabi ahaanba hoos u dhigaya 1µg ee pBR322DNA 10 daqiiqo gudahood at 37 ℃.
Xakamaynta tayada
RNase:5U of DNAse I oo leh 1.6µg MS2 RNA 4 saacadood oo ah 37℃ ma keenayso hoos u dhacWaxaa go'aamiya agarose gel electrophoresis.
Xusuusin
1. Fadlan diyaari 0.5MEDTA keligaa.
2. Isticmaal 1U DNAse I halkii µg ee RNA.Si kastaba ha noqotee, haddii RNA uu ka yar yahay 1µg, fadlan isticmaal 1U DNAse I.
3. Fadlan ku rid insaymiska dushiisa baraf xilliga shaqada.
