Rnase Inhibitor (Glycerol kubuntu)
Murine RNase inhibitor ni recombinant murine RNase inhibitor yagaragajwe kandi yeza kuri E.coli.Ihuza na RNase A, B cyangwa C mu kigereranyo cya 1: 1 binyuze mu guhuza kudahuza, bityo bikabuza ibikorwa byimisemburo itatu kandi bikarinda RNA kwangirika.Ariko, Ntabwo ari byiza kurwanya RNase 1, RNase T1, S1 Nuclease, RNase H cyangwa RNase kuva Aspergillus.Inhibitor ya Murine RNase yageragejwe na RT-PCR, RT-qPCR na IVT mRNA, kandi yari ihujwe n’ubucuruzi butandukanye bwa Reverse transcriptase, ADN polymerase na polymerase ya RNA.
Ugereranije na RNase inhibitor ya muntu, murine RNase inhibitor ntabwo irimo sisitemu ebyiri zumva cyane okiside itera kudakora kwa inhibitor.Ibyo bituma bihagarara neza kuri DTT (munsi ya mM 1).Iyi mikorere ituma ikwiriye gukoreshwa mubitekerezo aho kwibanda cyane kwa DTT bihabanye nigisubizo (urugero Real-time RT-PCR).
Agusaba
Ibicuruzwa birashobora gukoreshwa cyane mubigeragezo ibyo aribyo byose aho kwivanga kwa RNase bishoboka kugirango wirinde kwangirika kwa RNA, nka:
1.Icyiciro cya mbere cDNA synthesis, RT-PCR, RT-qPCR, nibindi.;
2.Kurinda RNA gutesha agaciro muri vitro transcription / ibisobanuro (urugero: kwigana virusi muri vitro);
3.Kubuza ibikorwa bya RNase mugihe cya RNA kwigunga no kwezwa.
Ububiko
Ubike kuri -25 ～ -15 ℃;
Inzinguzingo zikonjesha times inshuro 5;
Byemewe kumyaka 1.
Igisobanuro cyibice
Igice kimwe gisobanurwa nkubunini bwa RNase Inhibitor isabwa kugirango uhagarike ibikorwa bya 5 ng ya RNase A kuri 50%.
Uburemere bwa molekile
Inhibitori ya RNase (idafite Glycerol) ni proteine 50 kDa.
Kugenzura ubuziranenge
Exonuc Please Igikorwa:
Kwiyongera kwa 40 U ya enzyme hamwe na 1 μg λ-Hind III igogora ADN amasaha 16 kuri 37 ° C byatumye nta kwangirika kugaragara kwa ADN nkuko byagenwe na gel electrophorei.
Igikorwa cya Endonuc Please:
Kwiyongera kwa 40 U ya enzyme hamwe na 1 μg λ ADN mu masaha 16 kuri 37 ° C byatumye nta kwangirika kugaragara kwa ADN nkuko byagenwe na gel electrophorei.
Nicking Igikorwa:
Kwiyongera kwa 40 U ya enzyme hamwe na 1 μg pBR322 kumasaha 16 kuri 37 ℃ byatumye nta kwangirika kugaragara kwa ADN nkuko byagenwe na gel electrophorei.
RNase Igikorwa:
Inkubasi ya 40 U ya enzyme hamwe na 1,6 μg MS2 RNA mumasaha 4 kuri 37 ℃ byatumye nta kwangirika kugaragara kwa RNA nkuko byagenwe na gel electrophorei.
E.ADN ya coli:
40 U ya Enzyme igaragazwa na TaqMan qPCR.ADN ya E.coli ni ≤ 0. 1pg / 40U.
Notes
1.Ntugahungabanye cyangwa ngo ubyuke bikabije kugirango wirinde gukora enzyme.
2.RNase inhibitor ikora mubushyuhe buri hagati ya 25 ℃ kugeza 55 ℃ kandi idakora kuri ≥65 ℃.
3.Ibikorwa bya RNase H, RNase 1, na RNase T1 ntibibujijwe.
4.Urwego rwa pH rwo guhagarika ibikorwa bya RNase ni rugari (rukora kuri pH 5-9), rugaragaza ibikorwa byinshi kuri pH 7-8.
