Description
The ExcelTaq™ 5X/2X PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl2 (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel electrophoresis.
Features
5’→3’ DNA polymerase activity
No detectable 3'→5' exonuclease (proofreading) activity
Generates PCR products with 3'-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
Applications
Routine PCR
Colony PCR
High throughput PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
4°C for 6 months
-20°C for 24 months
Elongation capability
ExcelTaq™ PCR Master Mix can reliably amplify λDNA up to 8 kb in length. (M: DM3100)
Contents
Component |
Volume |
Cat. No |
ExcelTaq™ 5X PCR Master Mix |
1 ml x 2 |
TP1100 (200 Rxn) |
6X DNA Loading Dye (Blue) | 1 ml x 2 | |
ExcelTaq™ 2X PCR Master Mix (MgSO4) |
1.25 ml x 2 |
TP1120 (100 Rxn) |
6X DNA Loading Dye (Blue) |
1 ml |
Storage
4°C for 6 months
-20°C for 24 months
Recommended PCR Condition
|
Recommended PCR Program
Steps |
Temp. |
Time |
Cycles |
Template denature |
94°C |
2 min |
1 |
Denature |
94°C |
30 sec |
25-40 |
Annealing |
50-68°C* |
30 sec |
|
Extension |
72°C |
30 sec/kb |
|
Final extension |
72°C |
1 min |
1 |
*Optimal PCR condition varies according to primers’ thermodynamic properties.
Amin OM, Heckmann RA, Dallarés S, Constenla M, Ha NV.
Parasite. 2019;26:14. doi: 10.1051/parasite/2019015. Epub 2019 Mar 6.
PMCID: PMC6402367
High Fidelity PCR amplification
Amplification of target gene with HiFi™ DNA polymerase to minimize error rate.
Gel electrophoresis
Staining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.
Safe fluorescent dyes
[NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl
[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl
[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 ml
Blue-light illuminator
Ligation
Blund-end PCR amplicons can directly ligate with PCR cloning vector.
Transformation
Prepare competent cells with high efficiency and transform with time-saving protocol.