What is Ex Taq?
Ex Taq DNA polymerase—a robust PCR enzyme with proofreading activity. TaKaRa Ex Taq DNA Polymerase combines the proven performance of Takara Taq polymerase with the proofreading activity of an efficient 3′-to-5′ exonuclease, for high-sensitivity, high-efficiency PCR reactions.
How do I set PCR conditions?
A standard polymerase chain reaction (PCR) setup consists of four steps:
- Add required reagents or mastermix and template to PCR tubes.
- Mix and centrifuge.
- Amplify per thermo cycler and primer parameters.
- Evaluate amplified DNA by agarose gel electrophoresis followed by ethidium bromide staining.
Why is Taq used in PCR?
PCR amplification works on the principle of temperature variation—heating and cooling reactions—which makes Taq polymerase a highly advantageous enzyme. The major reason behind this is that Taq polymerase can work at high temperatures with high efficiency and amplification capacity, which other bodily enzymes cannot.
How much Taq do I add to PCR?
Taq DNA Polymerase Concentration: We generally recommend using Taq DNA Polymerase at a concentration of 25 units/ml (1.25 units/50 μl reaction). However, the optimal concentration of Taq DNA Polymerase may range from 5–50 units/ml (0.25–2.5 units/50 μl reaction) in specialized applications.
What are the advantages of Taq polymerase?
Thermostability: Taq polymerase is a thermostable DNA polymerase isolated from a bacterium that lives in hot springs. It can withstand the high temperature of >90°C required for the denaturing step in PCR and remain enzymatically active after each cycle.
What does Taq enzyme do?
Taq DNA polymerase is the most common enzyme used for PCR amplification. This enzyme is extremely heat resistant with a half-life of 40 minutes at 95°C. At its optimal temperature (72°C), nucleotides are incorporated at a rate of 2–4 kilobases per minute.
What is the role of Taq?
Taq polymerase is the heat-stable (thermostable) DNA polymerase extracted from the thermophilic bacteria Thermus aquaticus. Its predominant function is in the polymerase chain reaction (PCR) technique, where it automates the repetitive step of amplifying specific DNA sequences.
Why is Taq polymerase preferred in PCR?
The Role of Taq Polymerase in PCR Taq DNA Polymerase is highly efficient, so it becomes fully functional as it reaches its optimum temperature. It also has a half-life of more than two hours (at a temperature of 92 °C), a high-amplification capacity, and the ability to add 150 nucleotides per second.
Why did my PCR fail?
Forgetting just one component of the PCR reaction, whether that be the DNA polymerase, primers or even the template DNA, will result in a failed reaction. The simplest solution is to repeat the reaction. Take your time to ensure everything has been added.
What happens if you use too much Taq polymerase in PCR?
Too much Taq will result in an excessive background of unwanted DNA fragments (a smear on a gel) while a huge excess may cause the reaction to fail with no product being detected. A Taq concentration of 1 unit per 25μl reaction ensures a cleaner product and lower background.
What is the function of Taq in PCR?
Why only Taq polymerase is used in PCR?
Due to its key role in synthesizing and amplifying new strands of DNA, Taq DNA Polymerase is essential to Polymerase Chain Reaction (PCR). Like other DNA polymerases, Taq Polymerase can only produce DNA if it has a primer, a short sequence of 20 nucleotides that provide a starting point for DNA synthesis.
How do you know if PCR is successful?
Comparing your PCR samples to control samples (tubes not subjected to PCR) will confirm the success of PCR. Your PCR samples and control samples will be run alongside a DNA ladder. A DNA ladder contains DNA fragments of known size, measured in base pairs (bp).
What is Takara ex Taq DNA polymerase?
www.clontech.com/takara TaKaRa Ex Taq DNA Polymerase is a thermostable DNA polymerase having an efficient 3’–>5’ exonuclease activity. This for- mulation provides unsurpassed PCR performance, even for difficult amplifications.
Can Takara products be used for resale?
Also, do not use this product as food, cosmetic, or household item, etc. Takara products may not be resold or transferred, modified for resale or transfer, or used to manufacture commercial products without written approval from TAKARA BIO INC.
Is the Takara logo a trademark?
The Takara logo is a trademark of TAKARA HOLDINGS, Kyoto, Japan. All other marks are the property of their respective owners. Certain trademarks may not be registered in all jurisdictions. TAKARA BIO INC. 800-662-2566 www.clontech.com/takara