2 × HiF Taq plus Master Mix
Cat No: HCR2014B
HIF Taq ntxiv rau Master Mix (Nrog Dye) yog npaj-rau-siv 2 × premixed tov uas muaj Ntxiv HIF DNA Polymerase, dNTPs, thiab optimized buffer.Ob lub monoclonal antibodies nyob rau hauv chav tsev kub uas inhibit polymerase kev ua si thiab 3′→ 5′ exonuclease kev ua tau ntxiv rau tus tswv mix rau yooj yim thiab heev tshwj xeeb kub Start PCR.Qhov txuas ntxiv tau ntxiv rau tus tswv sib xyaw kom muab cov enzyme kom muaj peev xwm ua kom lub zog ntev ntev, qhov ntev ntawm kev nthuav dav tuaj yeem ncav cuag 13 kb, enzyme muaj 5' → 3 ' DNA polymerase kev ua haujlwm thiab 3' → 5' exonuclease kev ua, nws fidelity yog 83 npaug ntawm Taq DNA polymerase, uas yog 9 npaug ntawm zoo tib yam DNA polymerase.Nws yog tsim rau amplification ntawm complex templates, lub amplification khoom yog ib tug blunt kawg.
2 × HIF Taq ntxiv rau Master Mix (Nrog Dye) muaj qhov zoo ntawm kev ceev thiab yooj yim, siab rhiab heev, muaj zog tshwj xeeb, zoo stability, thiab lwm yam., cov tshuaj tiv thaiv tsuas yog yuav tsum tau ntxiv primers thiab templates, thiab yuav tsum tau amplified los ntawm ob- kauj ruam raws tu qauv, simplifying cov kauj ruam sim thiab txuag lub sij hawm.Cov khoom no muaj cov cim qhia electrophoresis, thiab cov khoom PCR tuaj yeem siv ncaj qha rau electrophoresis.Tsis tas li ntawd, cov khoom kuj tseem muaj cov neeg sawv cev tiv thaiv tshwj xeeb, kom tus tswv sib xyaw tuaj yeem tswj hwm kev ua haujlwm ruaj khov tom qab rov khov-thaw.
Kev cia khoom
Cov khoom yuav tsum khaws cia ntawm -25 ~ -15 ℃ rau 1 xyoos.
Specifications
| Khoom specification | Master Mix |
| Nco ntsoov | 2 × |
| Pib kub | Built-in Kub Pib |
| Overhang | Blunt |
| Cov tshuaj tiv thaiv ceev | Ceev ceev |
| Size (Cov khoom kawg) | Mus txog 13 kb |
| Cov xwm txheej rau kev thauj mus los | Dej khov qhuav |
| Yam khoom | High fidelity PCR premixes |
Cov lus qhia
1.PCR Reaction System
| Cheebtsam | Ntim (μL) |
| DNA Template | Tsim nyog |
| Forward primer (10 μmol / L) | 2.5 |
| Rov qab Primer (10 μmol / L) | 2.5 |
| 2 × HIF Taq plus Master Mix | 25 |
| ddH ua2O | rau 50 |
2.Pom zoo siv cov qauv sib txawv
| Hom qauv | Amplify fragments ntawm 1kb mus rau 10 kb |
| Genomic DNA | 50-200 hli |
| Plasmid lossis Viral DNA | 10 pg-20 ib |
| cDNA | 1-2.5 µL (Tsis pub tshaj 10% ntawm cov tshuaj tiv thaiv PCR zaum kawg) |
3.Amplification raws tu qauv
1) Ob-kauj ruam raws tu qauv (cov qauv nyuaj)
| Lub voj voog kauj ruam | Temp. | Sijhawm | Lub voj voog |
| Thawj denaturation | 98 ℃ | 3 feeb | 1 |
| Denaturation | 98 ℃ | 10 sec | 30-35 : kuv |
| Txuas ntxiv | 68 ℃ | 30 sec/kb | |
| Qhov kawg txuas ntxiv | 72 ℃ | 5 feeb | 1 |
2) Peb-kauj ruam raws tu qauv (cov txheej txheem tsis tu ncua)
| Lub voj voog kauj ruam | Temp. | Sijhawm | Lub voj voog |
| Thawj denaturation | 98 ℃ | 3 feeb | 1 |
| Denaturation | 98 ℃ | 10 sec | 30-35 : kuv |
| Annealing | 60 ℃ | 20 sec | |
| Txuas ntxiv | 72 ℃ | 30 sec/kb | |
| Qhov kawg txuas ntxiv | 72 ℃ | 5 feeb | 1 |
3) Annealing Gradient Protocol (cov qauv nyuaj)
| Lub voj voog kauj ruam | Kub | Sijhawm | Lub voj voog |
| Thawj denaturation | 98 ℃ | 3 feeb | 1 |
| Denaturation | 98 ℃ | 10 sec | 15 (1 ℃ txo ib lub voj voog) |
| Gradient annealing | 70-55 ℃ | 20 sec | |
| Txuas ntxiv | 72 ℃ | 30 sec/kb | |
| Denaturation | 98 ℃ | 10 sec |
20 |
| Annealing | 55 ℃ | 20 sec | |
| Txuas ntxiv | 72 ℃ | 30 sec/kb | |
| Qhov kawg txuas ntxiv | 72 ℃ | 5 feeb | 1 |
Nta nyob rau hauv txawv amplification raws tu qauv
| Protocol | Ob-kauj ruam | Peb-kauj ruam | Gradient annealing |
| Spec. | ceev | nruab nrab | qeeb |
| Qhov tshwj xeeb | siab | nruab nrab | siab |
| PCR tawm los | nruab nrab | siab | nruab nrab |
| Kev kuaj xyuas tus nqi | siab | nruab nrab | siab |
Sau ntawv
Thov hnav PPE tsim nyog, xws li lub tsho tiv no thiab hnab looj tes, txhawm rau ua kom koj txoj kev noj qab haus huv thiab kev nyab xeeb!
