SureStart Taq DNA Polymerase - Details & Specifications
SureStart Taq DNA polymerase is a modified version of Taq2000 DNA polymerase with hot start capability. SureStart Taq DNA polymerase improves PCR amplification reactions by decreasing background noise and increasing amplification of desired products. Using SureStart Taq, hot start is easily incorporated into PCR protocols already optimized with Taq DNA polymerase, with little or no modification of cycling parameters or reaction conditions.
Preparing PCR reaction mixtures at room temperature can result in high background and lower yields of specific products. Certain PCR enzymes exhibit significant polymerase activity at temperatures encountered during reaction setup or while ramping up to stringent primer annealing temperatures. For example, Taq DNA polymerase exhibits 2–3% maximum activity at 25°C (e.g., room temperature setup) and 70% maximum activity at 50°C (which is generally below the melting temperature of PCR primers). Nonspecific primer annealing and extension at nonrestrictive temperatures produces undesirable products that are amplified throughout the remaining cycles. Misprimed products and artifacts such as primer-dimers can impair gel analysis, quantitation, and sequencing of specific PCR products. In the amplification of misprimed products, dNTPs and primers are diverted away from the synthesis of specific products, reducing overall yields.
A number of hot start techniques have been developed to improve amplification specificity and to allow PCR setup at ambient temperatures. The most convenient hot start methods employ reversibly inactivated enzymes. With SureStart Taq DNA polymerase, hot start is provided by a modified form of Agilent's Taq2000 DNA polymerase, a highly purified recombinant version of Taq DNA polymerase. SureStart Taq DNA polymerase remains inactive until stringent temperatures (e.g., 92–95?C) are reached. SureStart Taq can be activated by adding an initial step of 9–12 minutes at 92–95°C to the beginning of PCR cycling programs. Alternatively, the enzyme can be activated slowly during temperature cycling without prior activation, although it may be necessary to add additional cycles to existing cycling programs to achieve optimal product yield. Either activation method provides a PCR hot start, since primer extension can not occur during PCR setup when SureStart Taq DNA polymerase is inactive. SureStart Taq can be used in a variety of amplification systems, including quantitative PCR and RT-PCR, to improve specificity, yield, and amplification of difficult targets.
SureStart Taq DNA polymerase is optimized for use in amplifying DNA targets between 100 and 2000 bases. Although templates of up to 5 kb may be amplified with SureStart Taq DNA polymerase, we recommend the use of Herculase enhanced DNA polymerase to achieve optimal yields of longer targets (>2 kb).
For Research Use Only. Not for use in diagnostic procedures