UDG/UNG Enzymes
UDG (uracil DNA glycosylase) tuaj yeem ua rau lub hydrolysis ntawm N-glycosidic txuas ntawm uracil puag thiab qab zib-phosphate qaum hauv ssDNA thiab dsDNA.Nws tuaj yeem tswj tau yooj yim aerosol muaj kuab paug thiab haum rau cov kab ke molecular biology xws li PCR, qPCR, RT-qPCR thiab LAMP.
Specifications
| Expression Host | Recombinant E. coliwith uracil DNA glycosylase noob |
| Molecular Luj | 24.8kd ua |
| Purity | ≥95% (SDS-PAGE) |
| Kub Inactivation | 95 ℃, 5 ~ 10 min |
| Unit txhais | Ib chav tsev (U) txhais tau tias yog tus nqi ntawm cov enzyme uas yuav tsum tau catalyze hydrolysis ntawm l μg dU-muaj dsDNA hauv 30 feeb ntawm 25 ℃. |
Cia
Cov khoom yuav tsum khaws cia ntawm -25 ℃ ~ -15 ℃ rau ob xyoos.
Cov lus qhia
1.Kev npaj ntawm PCR cov tshuaj tiv thaiv sib tov raws li hauv qab no
| Cheebtsam | Ntim (μL) | Thaum kawg concentration |
| 10 × PCR Buffer (Mg² + Ntxiv) | 5 | 1 × |
| 25 mmol/LMgCl | 3 | 1.5 mmol / L |
| dUTP (10 mmol / L) cov tsos mob | 3 | 0.6 mmol / L |
| dCTP/dGTP/dATP/dTTP (10mmol/Leach) | 1 | 0.2 mmol / L - ntshav qab zib |
| Template DNA | X | - |
| Primer 1 (10 μmol / L) | 2 | 0.4 μmol / L |
| Primer 2 (10 μmol / L) | 2 | 0.4 μmol / L |
| DNA Polymerase (5 U / μL) | 0.5 | 0.05 U/μL |
| Uracil DNA Glycosylase (UDG/UNG), 1 U/μL | 1 | 1 U / μL |
| ddH₂O | Txog li 50 | - |
Nco tseg: Raws li kev sim xav tau, qhov kawg ntawm dUTP tuaj yeem hloov kho ntawm 0.2-0.6 mmol / L, thiab 0.2 mmol / L dTTP tuaj yeem raug xaiv ntxiv.
2.Amplification txheej txheem
| Lub voj voog kauj ruam | Kub | Sijhawm | Lub voj voog |
| dU-muaj template degradation | 25 ℃ | 10 feeb | 1 |
| UDG qhib, template pib denaturation | 95 ℃ | 5 ~ 10 feeb | 1 |
| Denaturation | 95 ℃ | 10 sec |
30-35 : kuv |
| Annealing | 60 ℃ | 20 sec | |
| Txuas ntxiv | 72 ℃ | 30 sec/kb | |
| Qhov kawg txuas ntxiv | 72 ℃ | 5 feeb | 1 |
Nco tseg: Lub sijhawm tshuaj tiv thaiv ntawm 25 ° C tuaj yeem hloov kho tsis pub dhau 5-10mins raws li kev sim.
Sau ntawv
1.UDG ua haujlwm hauv feem ntau PCR cov tshuaj tiv thaiv buffers.
2.Cov enzymes yuav tsum muab cia rau hauv lub thawv dej khov lossis hauv lub da dej dej khov thaum siv, thiab yuav tsum khaws cia ntawm -20 ° C tam sim tom qab siv.
3.Thov hnav PPE tsim nyog, xws li lub tsho tiv no thiab hnab looj tes, txhawm rau ua kom koj noj qab haus huv thiab nyab xeeb!
