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While microbes are often viewed negatively due to their association with many human illnesses, microbes are also responsible for many beneficial processes such as industrial fermentation (e.g. the production of alcohol and dairy products), antibiotic production and as vehicles for cloning in higher organsisms such as plants. Scientists have also exploited their knowledge of microbes to produce biotechnologically important enzymes such as Taq polymerase, reporter genes for use in other genetic systems and novel molecular biology techniques such as the yeast two-hybrid system.
Taq polymerase ("Taq Pol," or simply "Taq") is a thermostable polymerase used in polymerase chain reaction to check for the presence or absence of a gene by amplifying a DNA fragment. First isolated from Thermus aquaticus, a bacterium that lives in hot springs and hydrothermal vents, Taq was identified as the first polymerase able to withstand the denaturing conditions required during PCR. Its enzymatic halflife (at 95 degrees Celsius) is 40 min.
One of Taq polymerases' major drawbacks is its low replication fidelity. Without a 3' to 5' exonuclease proofreading mechanism to replace an accidental mismatch in the newly synthesized DNA strand, Taq polymerase cannot be used in experiments where an identical genetic sequence is required (such as in molecular cloning). Commercially sold Taq DNA polymerase has an error rate of one in 10,000 nucleotides and typically produces 16% of mutated 1kb (kilobasepair) PCR products in a reaction. It can amplify a 1kb strand of DNA in roughly 30 seconds at 72°C.
Taq DNA polymerase also yields fragments with A (Adenine) overhangs. This is particularly useful in 'TA Cloning,' whereby a cloning vector (such as a plasmid) is used which has T (Thymine) overhangs which complement with the A overhangs of the PCR Product, thus increasing the ligation efficiency.
Pfu DNA polymerase is often used instead of, or in conjunction with, Taq polymerase. This enzyme is more thermostable and proofreads DNA, resulting in a lower error rate.
Leading commercial suppliers of Taq DNA Polymerase include [1]Promega, Roche and Invitrogen