What is GoTaq Mastermix?
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?
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).
What does magnesium do in PCR?
Magnesium ion (Mg2+) functions as a cofactor for activity of DNA polymerases by enabling incorporation of dNTPs during polymerization. The magnesium ions at the enzyme’s active site catalyze phosphodiester bond formation between the 3′-OH of a primer and the phosphate group of a dNTP (Figure 6).
Why is GoTaq master mix dyed?
GoTaq® Green Master Mix contains two dyes (blue and yellow) that allow monitoring of progress during electrophoresis.
What is in a master mix for PCR?
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.
What is the purpose of colony PCR?
Colony PCR is a method for rapidly screening colonies of yeast or bacteria that have grown up on selective media following a transformation step, to verify that the desired genetic construct is present, or to amplify a portion of the construct.
What is green Taq?
Description. Green Taq DNA Polymerase is designed to increase the stability of the Taq enzyme for more convenient transport and applications. The polymerase (1000 U) is designed for 400 rxns if 2.5 U are used per 50 ul PCR reaction.
What is the purpose of adding mg2+ in a PCR reaction?
The Role of MgCl2 in PCR reaction is to enhance the DNA amplification by boosting the activity of Taq DNA polymerase. The polymerase chain reaction is one of the important experimental procedure in downstream genomic and genetic research. It generates millions of amplicon for the gene of our interest.
What enzymes are required for PCR?
Taq polymerase Like DNA replication in an organism, PCR requires a DNA polymerase enzyme that makes new strands of DNA, using existing strands as templates. The DNA polymerase typically used in PCR is called Taq polymerase, after the heat-tolerant bacterium from which it was isolated (Thermus aquaticus).
Does Taq polymerase have exonuclease activity?
Taq DNA polymerase has a domain at its amino terminus (residue 1 to 291) that has a 5′-3′ exonuclease activity, a 3′-5′ exonuclease domain in the middle (residue 292 to 423), and a domain at its C-terminus that catalyzes polymerase reactions. coli DNA polymerase I.
What are the 3 types of PCR?
Types of PCR
- Real-time PCR.
- Quantitative real time PCR (Q-RT PCR)
- Reverse Transcriptase PCR (RT-PCR)
- Multiplex PCR.
- Nested PCR.
- Long-range PCR.
- Single-cell PCR.
- Fast-cycling PCR.
Can you store PCR master mix?
In order to avoid the time-consuming process of thawing it is also possible to store all Ampliqon DNA polymerases, master mixes, buffers and PCR reagents at +4 °C for up to 6 months. RealQ Plus master mixes can be kept at + 4°C for up to 3 months. We strongly recommend shipping on dry ice to ensure maximal shelf life.
What is the protocol for GoTaq green master mix?
GoTaq® Green Master Mix Protocol. GoTaq® Green Master Mix is a premixed ready-to-use solution containing bacterially derived Taq DNA polymerase, dNTPs, MgCl 2 and reaction buffers at optimal concentrations for efficient amplification of DNA templates by PCR.
What is the concentration of MgCl 2 in GoTaq?
The 5X GoTaq® Green and Colorless Reaction Buffers supplied with GoTaq® DNA Polymerase contain MgCl 2 at a concentration of 7.5mM for a final concentration of 1.5mM in the 1X reaction. The 5X Green and 5X Colorless Reaction Buffers allow you to go directly from thermal cycler to gel analysis.
What kind of dyes are in GoTaq Green?
GoTaq® Green Master Mix also includes two dyes (blue and yellow) that allow monitoring of progress during electrophoresis. The blue dye comigrates at the same rate as 3–5kb DNA fragments in a 1% agarose gel.
What kind of PCR is used for GoTaq?
Use the green reaction buffer for direct-to-gel analysis after amplification and the colorless reaction buffer for post-amplification analysis by fluorescence or absorbance without prior DNA purification. PCR enhancers, such as betaine and DMSO, have no effect on the color or characteristics of the GoTaq® Green Reaction Buffer.