2 × PCR Super Mix (nrog Dye)
2 × PCR Master Mix muaj Taq DNA Polymerase, dNTPs, thiab lwm yam PCR-yuav tsum tau ua.Master Mix yog ruaj khov rau 3 lub hlis ntawm 4 ℃ nrog peb cov stabilizers customized.Cov tshuaj ua ntej sib xyaw yog ua kom zoo rau cov pa PCR thiab npaj siv los ntawm kev ntxiv DNA template thiab primers.Cov khoom PCR tuaj yeem thauj khoom ncaj qha rau electrophoresis nrog pre-loaded bromophenol xiav dye.Cov khoom lag luam amplified muaj 3 '-dA protrusion thiab tuaj yeem yooj yim cloned rau hauv T vector.2 × PCR Master Mix ua kom yooj yim rau PCR txheej txheem thiab txo cov kab mob.
Kev cia khoom
Cov khoom yuav tsum khaws cia ntawm -25 ℃ ~ -15 ℃ rau 2 xyoos.
Specifications
| Fidelity(vs.Taq) | 1 × |
| Pib kub | No |
| Overhang | 3 '-A |
| Polymerase | Cov DNA Polymerase |
| Hom tshuaj tiv thaiv | SuperMix lossis Master Mix |
| Teb ceev | Txuj |
| Yam khoom | PCR Master Mix (2x) |
Cov lus qhia
1.Reaction System
| Cheebtsam | Ntim(μL) |
| Template DNA | Tsim nyog |
| Primer 1 (10 μmol / L) | 2 |
| Primer 2 (10 μmol / L) | 2 |
| 2 × PCR Master Mix | 25 |
| ddH ua2O | rau 50 |
2.Amplification raws tu qauv
| Lub voj voog kauj ruam | Qhov kub thiab txias (°C) | Sijhawm | Lub voj voog |
| Thawj denaturation | 94 | 5 feeb | 1 |
| Denaturation | 94 | 30 sec | 35 |
| Annealing | 50-60 | 30 sec | |
| Txuas ntxiv | 72 | 30-60 sec/kb | |
| Qhov kawg txuas ntxiv | 72 | 10 feeb | 1 |
Nco tseg:
1) Cov qauv siv: 50-200ng genomic DNA;0.1-10ng plasmid DNA.
2) Mg2+concentration: Cov khoom no muaj 3mM ntawm MgCl2, haum rau feem ntau cov tshuaj tiv thaiv PCR.
3) Annealing kub: Thov xa mus rau qhov theoretical Tm tus nqi ntawm primers.Lub annealing kub tuaj yeem teem rau 2-5 ℃ qis dua tus nqi theoretical ntawm primer.
4) Lub sijhawm ncua sijhawm: Rau kev txheeb xyuas molecular, 30 sec / kb raug pom zoo.Rau cov noob cloning, 60 sec / kb raug pom zoo.
Sau ntawv
1.PCR khoom nrog 2 × PCR Master Mix tsis haum rau polyacrylamide gel electrophoresis.
2.Rau koj txoj kev nyab xeeb thiab kev noj qab haus huv, thov hnav lub tsho (lab) thiab cov hnab looj tes pov tseg rau kev ua haujlwm.
3.Nws yog siv rau kev tshawb fawb siv nkaus xwb!
