Rnase Inhibitor (Glycerol dawb)
Murine RNase inhibitor yog recombinant murine RNase inhibitor qhia thiab purified los ntawm E.coli.Nws khi rau RNase A, B lossis C hauv qhov sib piv 1: 1 los ntawm kev sib txuas tsis yog covalent, yog li inhibiting kev ua haujlwm ntawm peb enzymes thiab tiv thaiv RNA los ntawm degradation.Txawm li cas los xij, Nws tsis zoo rau RNase 1, RNase T1, S1 Nuclease, RNase H lossis RNase los ntawm Aspergillus.Murine RNase inhibitor tau kuaj los ntawm RT-PCR, RT-qPCR thiab IVT mRNA, thiab tau sib xws nrog ntau yam kev lag luam rov qab transcriptases, DNA polymerases thiab RNA polymerases.
Piv rau tib neeg RNase inhibitors, murine RNase inhibitor tsis muaj ob lub cysteine rhiab heev rau oxidation uas ua rau inactivation ntawm inhibitor.Qhov ntawd ua rau nws ruaj khov ntawm qhov tsis tshua muaj siab ntawm DTT (tsawg dua 1 mM).Cov yam ntxwv no ua rau nws tsim nyog rau kev siv cov tshuaj tiv thaiv uas muaj qhov siab ntawm DTT tsis zoo rau cov tshuaj tiv thaiv (xws li Real-time RT-PCR).
Adaim ntawv thov
Cov khoom no tuaj yeem siv dav hauv txhua qhov kev sim uas RNase cuam tshuam tau kom tsis txhob RNA degradation, xws li:
1.First-strand cDNA synthesis, RT-PCR, RT-qPCR, thiab lwm yam;
2. Tiv thaiv RNA los ntawm degradation nyob rau hauv-vitro transcription/translation (xws li viral replication nyob rau hauv vitro);
3.Inhibition ntawm RNase kev ua si thaum lub sij hawm RNA cais thiab purification.
Kev cia khoom
Khaws ntawm -25 ～ -15 ℃;
Lub voj voog khov-thaw ≤ 5 zaug;
Siv tau rau 1 xyoos.
Unit txhais
Ib chav tsev yog txhais raws li tus nqi ntawm RNase Inhibitor yuav tsum tau inhibit qhov kev ua ntawm 5 ng ntawm RNase A los ntawm 50%.
Molecular hnyav
RNase Inhibitor (Glycerol-dawb) yog 50 kDa protein.
Kev tswj kom zoo
Exonuclease Kev ua:
Incubation ntawm 40 U ntawm enzyme nrog 1 μg λ-Hind III zom DNA rau 16 teev ntawm 37 ° C ua rau tsis pom qhov degradation ntawm DNA raws li kev txiav txim los ntawm gel electrophoresis.
Kev Ua Haujlwm Endonuclease:
Incubation ntawm 40 U ntawm enzyme nrog 1 μg λ DNA rau 16 teev ntawm 37 ° C ua rau tsis pom qhov degradation ntawm DNA raws li kev txiav txim los ntawm gel electrophoresis.
Nicking Kev ua:
Incubation ntawm 40 U ntawm enzyme nrog 1 μg pBR322 rau 16 teev ntawm 37 ℃ ua rau tsis pom qhov degradation ntawm DNA raws li kev txiav txim los ntawm gel electrophoresis.
RNase Kev ua si:
Incubation ntawm 40 U ntawm enzyme nrog 1.6 μg MS2 RNA rau 4 teev ntawm 37 ℃ ua rau tsis pom qhov degradation ntawm RNA raws li kev txiav txim los ntawm gel electrophoresis.
E.coli DNA:
40 U ntawm Enzyme raug kuaj pom los ntawm TaqMan qPCR.E.coli DNA yog ≤ 0. 1pg/40U.
Nua otes
1.Tsis txhob co los yog do nruj nreem kom tiv thaiv enzyme inactivation.
2.RNase inhibitor yog ua haujlwm ntawm qhov kub ntawm 25 ℃ mus rau 55 ℃ thiab yog inactivated ntawm ≥65 ℃.
3.Qhov kev ua ntawm RNase H, RNase 1, thiab RNase T1 tsis inhibited.
4.Qhov pH ntau rau inhibiting RNase kev ua haujlwm yog dav (ua haujlwm ntawm pH 5-9), nthuav tawm cov haujlwm siab tshaj plaws ntawm pH 7-8.
