DNA I
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 ku xiran tahay Ca2+ waxaana lagu kicin karaa ion birta kala duwan sida Mn2+ iyo Zn2+.5mM Ca2+ waxay ka ilaalisaa enzyme-ka hydrolysis.Joogitaanka Mg2+, ensaymku wuxuu si aan kala sooc lahayn u aqoonsan karaa oo u kala goyn karaa meel kasta oo ku taal qayb kasta oo DNA ah.Joogitaanka Mn2+, labada qaybood ee DNA-da ayaa isku mar la aqoonsan karaa oo la kala jeexayaa isla goobta si ay u sameeyaan jajabyo DNA-da oo siman ama jajab dhammaadka dhegdhegaysan ee DNA oo leh 1-2 nucleotides.
Hantida Alaabta
Bovine Pancreas DNAse I waxaa lagu muujiyay habka muujinta khamiirka waana la nadiifiyay.
Cxoogsheegayaal
| Qayb | Mugga | |||
| 0.1KU | 1KU | 5KU | 50KU | |
| DNAse I, RNase-free | 20μL | 200μL | 1ml | 10ml |
| 10×DNase I Buffer | 1ml | 1ml | 5× 1ml | 5× 10ml |
Gaadiidka iyo Kaydinta
1. Dejinta Kaydinta: - 15 ℃ ~ -25 ℃ kaydinta;
2.Transport Stability: Gaadiidka hoos baakadaha barafka;
3. Waxaa lagu keenay: 10 mM Tris-HCl, 2 mM CaCl2, 50% glycerol, pH 7.6 at 25 ℃.
Qeexitaan Unug
Hal unug waxa lagu qeexaa cadadka insaymka kaas oo si buuxda hoos u dhigaya 1 μg ee pBR322 DNA 10 daqiiqo gudahood 37°C.
Xakamaynta tayada
RNase:5U of DNase I oo leh 1.6 μg MS2 RNA 4 saacadood oo ah 37 ℃ ma keenayso hoos u dhac sida lagu go'aamiyay agarose gel electrophoresis.
Bakteeriya Endotoxin:LAL-tijaabo, marka loo eego daabacaadda Chinese Pharmacopoeia IV 2020, habka tijaabada xadka jel, qaanuunka guud (1143).Bakteeriyada endotoxin waa inay noqotaa ≤10 EU/mg.
Tilmaamaha Isticmaalka
1. Ku diyaari xalka falcelinta tuubada-free RNase sida waafaqsan saamiga hoos ku taxan:
| Qayb | Mugga |
| RNA | X μg |
| 10 × DNase I Buffer | 1 μL |
| DNA-ga I, RNase-free (5U/μL) | 1 U halkii μg RNA |
| ddH2O | Ilaa 10 μl |
2.37 ℃ 15 daqiiqo;
3.Ku dar bakhaarka joojinta si aad u joojiso falcelinta, oo kuleyl 65℃ ilaa 10 daqiiqo si aad u hawlgeliso DNA-da I. Muunada waxaa si toos ah loogu isticmaali karaa tijaabada soo-qaadista ee xigta.
Xusuusin
1. Isticmaal 1U DNAse I halkii μg ee RNA, ama 1U DNAse I wax ka yar 1μg ee RNA.
2.EDTA waa in lagu daraa fiirsashada kama dambaysta ah ee 5 mM si looga ilaaliyo RNA inay hoos u dhigto inta lagu jiro firfircoonida enzyme.
