5x PCR-MM "ready-to-load"
5x PCR Master Mix “ready-to-load” is an optimized, ready-to-use solution for precise and reliable polymerase chain reactions (PCR) followed by visualization via agarose gel electrophoresis. The master mix is 5x concentrated and contains all components for efficient PCR (thermostable Taq DNA polymerase, reaction buffer, MgCl2, and dNTPs) in optimal concentrations (5x). All you need to do is add your desired template DNA, primers, and water, and you can start DNA amplification immediately.
The 5x PCR Mastermix “ready to load” contains blue and yellow dyes for identifying DNA fragments via agarose gel electrophoresis. The master mix can be used directly and conveniently for loading the agarose gel (“ready-to-load”).
Note: This master mix contains Tween-20. If a detergent-free buffer system is required, please contact us.
Properties
- 5x concentrated, ready-to-use master mix
- Thermostable Taq DNA polymerase (Stable for up to one month at room temperature!)
- 12.5 mM MgCl₂ (= 2.5 mM in 1x solution)
- 1 mM each of dATP, dGTP, dCTP, and dTTP (= 200 µM each of dNTP in 1x solution)
- Yellow + blue dye for visualization via agarose gel electrophoresis
Good to know
How do the dyes behave in the agarose gel?
In a 1% agarose gel, the blue dye migrates at the same speed as DNA fragments ranging from 3.5 to 4.5 kb. In contrast, the yellow dye migrates at approximately the same speed as the primers (< 35–45 bp).
Do not use our 5x “ready-to-load” PCR master mix for applications involving subsequent spectrophotometric measurements (absorbance or fluorescence measurements), as the two dyes may interfere with these applications.
Which Taq DNA polymerase is used in the master mix?
Our optimized Taq DNA polymerase (isolated from the thermophilic bacterium Thermus aquaticus) was recombinantly expressed in E. coli and is characterized by high thermal stability. It remains stable at room temperature for up to 1 month without loss of activity (or for up to 6 months at 2–8 °C). Bio&SELL’s Taq DNA polymerase is a highly processive 5’ -> 3’ DNA polymerase with 5’ -> 3’ exonuclease activity. The enzyme adds single nucleotides (almost exclusively adenosine) to the 3’ ends of the DNA, making TA cloning possible without further modification.