Description
The ExcelTaq™ 5X PCR Master Dye Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as ready-to-use master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of template and primers. This not only saves valuable time in the laboratory, but also reduces the number of pipetting and reagent handling errors. The PCR Master Dye Mix is supplied as a 5X concentrated ready-to-use mix, that is a mixture of recombinant Taq DNA polymerase, reaction buffer, MgCl2, dNTP, enzyme stabilizer and PCR-friendly loading dye solution containing tracking dye (Bromophenol blue) enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent – loading dye master mix conveniently.
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
Includes tracking dye for direct loading after PCR
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 Dye Mix | 1 ml x 2 | TP1200 (200 Rxn) |
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.
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.
P. Aprilia, R. Ummami, C.M. Airin, F. Aziz, P. Astuti J. Food Qual. Hazards Control 2022, 9(2): 112-117 doi: 10.18502/jfqhc.9.2.10648