Kub Sensitive UNG
Temperature Sensitive UNG (TS-UNG) yog ib qho tau los ntawm cov lus qhia recombinant hauv E. coli.Lub enzyme catalyzes tso tawm dawb uracil los ntawm uracil-muaj ib leeg- thiab ob-stranded DNA thiab tsis muaj zog tiv thaiv RNA.Piv nrog rau cov pa UNG enzyme ntawm E. coli noob keeb kwm, TS-UNG enzyme muaj kev ua haujlwm siab dua ntawm qhov kub thiab txias (20 ℃ ~ 37 ℃) thiab kub rhiab heev thiab yooj yim inactivated (50 ℃), tsis txhob degradation ntawm dUTP-muaj amplification. cov khoom ntawm chav tsev kub los ntawm cov khoom seem uas yuav nyob tom qab inactivation ntawm cov pa UNG enzyme.Yog li ntawd, TS-UNG enzyme tsis yog tsuas yog haum rau PCR kev tiv thaiv kev tiv thaiv kab mob, tab sis kuj zoo sib xws nrog RT-PCR amplification program thiab tuaj yeem siv rau hauv RT-PCR kev tiv thaiv kev sib kis.
Pom zoo daim ntawv thov
Kev Tiv Thaiv Kev Tiv Thaiv Kev Tshaj Tawm
Kev cia khoom
-20 ° C rau lub sij hawm ntev cia, yuav tsum tau tov zoo ua ntej siv, tsis txhob nquag khov-thaw.
Cia tsis
20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, Stabilizer, 50% Glycerol.
Unit txhais
Tus nqi ntawm enzyme yuav tsum degrade 1µg ntawm ib leeg-stranded DNA uas muaj dU bases nyob rau hauv 1 teev ntawm 37 ° C yog 1 units ntawm kev ua (U).
Kev Tswj Xyuas Zoo
1.SDS-PAGE electrophoretic purity ntau dua 98%
2.Kev ua haujlwm degradation, batch-rau-batch tswj, stability
3.Tsis muaj exogenous nuclease kev ua, tsis muaj exogenous endonuclease lossis exonuclease paug.
Cov lus qhia
| Cheebtsam | Ntim (μL) | Thaum kawg concentration |
| 10 × PCR Buffer (dNTP dawb, Mg²+dawb) | 5 | 1 × |
| dUTPs (dCTP, dGTP, dATP) | - | 200 μM |
| dUTP (hloov dTTP) | - | 200-600 hli |
| 25 mMgcl2 | 2 - 8 L | 1-4 mM |
| 5 U / μL Taq | 0.25 | 1.25 ua |
| 1 U/μLTS-UNG | 0.5 (0.1-0.5 hli) | 0.5U (0.1-0.5U) |
| 25 × Primer Mixib | 2 | 1 × |
| Template | - | ＜1μg / tshuaj |
| ddH₂O | Rau 50 | - |
Lus Cim: a: Yog tias siv rau qPCR/qRT-PCR, lub fluorescent sojntsuam yuav tsum muab ntxiv rau hauv cov tshuaj tiv thaiv.Feem ntau, qhov kawg primer concentration ntawm 0.2 μM tuaj yeem muab cov txiaj ntsig zoo;Thaum cov tshuaj tiv thaiv kev ua haujlwm tsis zoo, cov primer concentration tuaj yeem hloov kho hauv thaj tsam ntawm 0.2-1 μM.Feem ntau, kev sojntsuam concentration yog optimized nyob rau hauv thaj tsam ntawm 0.1-0.3 μM.Concentration gradient thwmsim tuaj yeem ua tau los nrhiav qhov zoo tshaj plaws ua ke ntawm primer thiab sojntsuam.
Sau ntawv
1.Qhov ntsuas kub zoo tshaj plaws ntawm TS-UNG enzyme yog qhov qis, thiab nws tuaj yeem ua kom zoo nyob rau hauv thaj tsam ntawm 20 ℃ ~ 37 ℃, qhov ntau npaum ntawm cov enzyme thiab lub sijhawm tshuaj tiv thaiv tuaj yeem ua kom zoo nyob rau hauv thaj tsam ntawm 0.1 ~ 0.5 U, 5 ~ 10 feeb;thiab cov enzyme tuaj yeem ua rau tsis ua haujlwm hauv cov txheej txheem rov qab hloov pauv.
2.Nws yog tsim rau PCR thiab RT-PCR los tiv thaiv kev sib kis.
3.Tsis txhob nquag khov-thaw, thiab tsis txhob cuam tshuam rau qhov kub hloov pauv loj.
4.Cov noob sib txawv yuav tsum tau nthuav dav muaj qhov sib txawv ntawm kev siv dUTP thiab rhiab heev rau UNG enzyme, yog li ntawd, yog tias kev siv UNG system ua rau txo qis hauv kev tshawb pom, cov tshuaj tiv thaiv yuav tsum tau kho thiab kho kom zoo, yog tias koj xav tau kev txhawb nqa, thov hu rau peb lub tuam txhab.
