Enzymes on ZAGENO
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The basics of enzymes
Enzymes lower the activation energy of a reaction, thus accelerating the reaction rate of the conversion of substrate to product. The increased rate can be by a factor of several million and without this increased reaction rate, life would not be able to be sustained here on Earth.
Most enzymes are proteins, but there are a few exceptions where RNA molecules can act as catalysts for reactions.
Since the enzyme is a catalyst, it is not consumed during the reaction. Thus, decreased reaction rates observed over time is not due to the enzyme concentration decreasing, but a combination of reduced availability of substrates and an accumulation of product.
Enzymes often work within very narrow pH and temperature ranges. Hence, it is essential that you use the correct buffers and optimal temperature ranges given in the protocol that is supplied with the enzyme.
Enzymes can lower the activation energy (ΔG‡, Gibbs free energy) of a reaction in several ways.
Stabilization of the transition state
Providing alternative pathways
Destabilization of the substrate ground state
The nomenclature of enzymes is determined by the substrate that is used in the reaction and with the addition of the suffix -ase. One example of this would be the alcohol-metabolizing enzyme alcohol dehydrogenase.
Generally, enzymes are divided into the following groups:
- Oxidoreductases: catalyze oxidation/reduction reactions
- Ligases: can join molecules with covalent bonds
- Transferases: can transfer functional groups, such as polymerases
- Isomerases: can catalyze isomerization changes
- Hydrolases: catalyze the hydrolysis of various bonds, including restriction enzymes
- Lyases: cleave various bonds (not hydrolysis and oxidation)
Isozymes is the name given to enzymes that are different, but are catalyzing the same chemical reaction.
Our Restriction Enzyme Troubleshoot is useful for solving any potential problems.
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For example, this comparison shows:
that the IPROOF™ HIGH-FIDELITY DNA POLYMERASE Kit from Bio-Rad accepts amplicons up to 37 kb.
that the TAQ PLUS DNA POLYMERASE Kit from G-Biosciences is the cheapest per unit.
that the PLATINUM™ SUPERFI™ GREEN DNA POLYMERASE Kit from Invitrogen is compatible with the widest range of templates.
This comparison clearly exemplifies how every kit has strengths and weaknesses. Depending on what features you require for your experiment, these details should enable you to make an informed decision on the right kit for you.
The ZAGENO comparison does not highlight one kit to be better than the other, as the kit of choice may vary between researchers - depending on each individual's preferred attributes. The best kit is the one that meets your needs - ZAGENO allows you to make an informed decision with minimum effort.
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