Why is GoTaq master mix dyed?
GoTaq® Green Master Mix contains two dyes (blue and yellow) that allow monitoring of progress during electrophoresis. The master mix is not recommended if any downstream applications use absorbance or fluorescence excitation, as the yellow and blue dyes in the reaction buffer may interfere with these applications.
What are the components of the GoTaq Green PCR Master Mix?
Description: GoTaq® Green Master Mix is a premixed ready-to-use solution containing bacterially derived Taq DNA polymerase, dNTPs, MgCl2 and reaction buffers at optimal concentrations for efficient amplification of DNA templates by PCR.
What is GoTaq polymerase?
GoTaq® DNA Polymerase is a proprietary formulation of Taq polymerase that gives robust amplification equal to and, in some cases, superior to that of standard Taq. The blue dye comigrates with 3–5kb DNA fragments in a 1% agarose gel. The yellow dye migrates ahead of primers (<50bp).
Why do we need buffer in the master mix?
What is the master mix and why do you need each component? It contains all the components for PCR mix to occur; including the individual building blocks of DNA (nucleotides, or dNTP’s), a special buffer to maintain optimum pH, salts, and MgCl2.
What should be annealing temperature in PCR?
The annealing temperature (typically between 48-72°C) is related to the melting temperature (Tm) of the primers and must be determined for each primer pair used in PCR. During the extension step (typically 68-72°C) the polymerase extends the primer to form a nascent DNA strand.
What is the function of Taq buffer in PCR?
“The function of Taq DNA polymerase in PCR is to amplify or synthesize DNA or gene of interest for various downstream applications. It’s a type of thermostable DNA polymerase, work at a higher temperature as well.”
What are the 4 major components of PCR master mixes?
The master mix usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Using a master mix reduces pipetting and risk of contamination, is convenient, saves time and preempts possible errors in mixing, making it ideal for high-throughput applications.
Why do we use buffer in PCR?
PCR buffer is necessary to create optimal conditions for activity of Taq DNA polymerase. Buffers often contain Tris-Hcl, KCl, and sometimes MgCl2. PCR Page 4 Polymerase Chain Reaction, 12/2004 4 buffers are often available in 10X concentration and are sometimes Taq formulation-specific.