2 × Rapid Taq Super Mix
Cat No: HCR2016A
2 × Rapid Taq Super Mix yog raws li kev hloov kho Taq DNA Polymerase, ntxiv qhov muaj zog txuas ntxiv, kev ua kom muaj zog ntxiv thiab ua kom zoo dua qhov tsis zoo, nrog rau kev ua haujlwm siab loj.Lub amplification ceev ntawm complex templates xws li genome nyob rau hauv 3 kb mus txog 1-3 sec / kb, thiab hais tias ntawm yooj yim templates xws li plasmids nyob rau hauv 5 kb mus txog 1 sec / kb.Cov khoom no tuaj yeem txuag tau PCR cov tshuaj tiv thaiv lub sijhawm.Nyob rau tib lub sijhawm, sib xyaw muaj dNTP thiab Mg2+, uas tuaj yeem nthuav dav tsuas yog los ntawm kev ntxiv cov primers thiab cov qauv, uas kuj ua rau kev ua haujlwm yooj yim ntawm cov kev sim.Tsis tas li ntawd, sib tov muaj electrophoretic qhia dye, uas tuaj yeem ncaj qha electrophoresis tom qab cov tshuaj tiv thaiv.Tus neeg sawv cev tiv thaiv hauv cov khoom no ua rau cov khoom sib xyaw ua haujlwm ruaj khov tom qab rov khov thiab thawing.Lub 3'-kawg band A ntawm cov khoom PCR tuaj yeem yooj yim cloned rau hauv T vector.
Cheebtsam
2 × Rapid Taq Super Mix
Kev cia khoom
PCR Master Mix khoom yuav tsum khaws cia ntawm -25 ~ -15 ℃ rau 2 xyoos.
Specifications
| Khoom specification | Rapid Taq Super Mix |
| Nco ntsoov | 2 × |
| Pib kub | Built-in Kub Pib |
| Overhang | 3'-A |
| Cov tshuaj tiv thaiv ceev | Ceev ceev |
| Size (Cov khoom kawg) | Mus txog 15 kb |
| Cov xwm txheej rau kev thauj mus los | Dej khov qhuav |
Cov lus qhia
1. Cov tshuaj tiv thaiv kab mob (50 μL)
| Cheebtsam | Loj (μL) |
| Qauv DNA * | haum |
| Forward primer (10 μmol / L) | 2.5 |
| Rov qab primer (10 μmol / L) | 2.5 |
| 2 × Rapid Taq Super Mix | 25 |
| ddH2 ua | rau 50 |
2.Amplification raws tu qauv
| Lub voj voog kauj ruam | Qhov kub thiab txias (°C) | Sijhawm | Lub voj voog |
| Predenaturation | 94 | 3 feeb | 1 |
| Denaturation | 94 | 10 sec |
28-35 : kuv |
| Annealing | 60 | 20 sec | |
| Txuas ntxiv | 72 | 1-10 sec/kb |
Pom zoo siv cov qauv sib txawv:
| Hom qauv | Segment siv ntau yam (50 μL cov tshuaj tiv thaiv kab mob) |
| Genomic DNA los yog E. coli kua | 10-1,000 Nws |
| Plasmid lossis kab mob DNA | 0.5-50 hli |
| cDNA | 1-5 µL (tsis pub ntau tshaj 1/10 ntawm tag nrho cov ntim ntawm cov tshuaj tiv thaiv PCR) |
| Pom zoo siv cov qauv sib txawv | |
Nco tseg:
1.Reagent siv: tag nrho thaw thiab sib tov ua ntej siv.
2. Annealing kub: Lub annealing kub yog universal Tm tus nqi, thiab tseem tuaj yeem teem 1-2 ℃ qis dua tus nqi primer Tm.
3. Kev nrawm nrawm: Teem 1 sec / kb rau cov qauv nyuaj xws li genome thiab E. coli hauv 1 kb;teeb 3 sec/kb rau cov qauv nyuaj xws li 1-3 kb genome thiab E. coli;teem 10 sec/kb rau cov qauv nyuaj tshaj 3 kb genome thiab E. coli.Koj tuaj yeem teeb tus nqi rau 1 sec / kb rau cov qauv yooj yim xws li plasmid tsawg dua 5 kb, 5 sec / kb rau cov qauv yooj yim xws li plasmid ntawm 5 thiab 10 kb, thiab 10 sec / kb rau cov qauv yooj yim xws li plasmid loj dua 10 kb.
Sau ntawv
1. Rau koj txoj kev nyab xeeb thiab kev noj qab haus huv, thov hnav cov tsho hauv chav kuaj thiab cov hnab looj tes pov tseg rau kev ua haujlwm.
2. Cov khoom no yog siv rau kev tshawb fawb nkaus xwb!
