Application
FastStart Taq DNA Polymerase is a thermostable, chemically modified form of recombinant Taq DNA Polymerase. This enzyme delivers superior results thanks to its unique enzyme design and optimized buffer system. FastStart Taq DNA Polymerase is inactive at temperatures below 75°C, but is activated by a 2- to 4-minute incubation step at 95°C.
PCR
Hot Start PCR
RT-PCR
Quality
Each lot of FastStart Taq DNA Polymerase is function-tested using serially diluted samples of human genomic DNA as starting templates. A 365-bp fragment from the human tPA gene (single copy gene) was amplified using specific tPA primers. After ethidium-bromide staining, a 365-bp PCR fragment can be reproducibly detected on a 1% agarose gel when starting with 50 pg of human genomic DNA and 40 cycles of amplification.
Furthermore, a PCR assay is performed (under standard conditions using the GC-RICH solution) on 200 ng human genomic DNA with primers specific for a 284-bp fragment of the Apo E gene (74% GC content). After 30 cycles, a PCR product is detectable as a single specific band.
Each lot of FastStart Taq DNA Polymerase is tested for the absence of contaminating activities such as exo- and endonucleases and nicking activity.
Contents
FastStart Taq DNA Polymerase, 5 U/µl in storage and dilution buffer
PCR Reaction Buffer, 10x conc. with 20 mM MgCl2
PCR Reaction Buffer, 10x conc. without MgCl2
MgCl2 Stock Solution, 25 mM
GC-RICH Solution, 5x conc.
Product Description
The enzyme dNTPack comprises FastStart Taq DNA Polymerase and PCR-Grade Nucleotides in a ready-to-use solution. FastStart Taq DNA Polymerase is a versatile enzyme that can be used in a wide variety of applications and on multiple instrument platforms. This modified recombinant Taq DNA Polymerase is inactive at temperatures below 75°C, but is activated by a 2- to 4-minute heat activation step at 95°C. An optimized PCR buffer system and a GC-RICH Solution to enable the enzyme to handle a wide range of templates are also supplied.
Background Information
Hot start protocols have been shown to significantly improve the specificity, sensitivity, and yield of PCR. Heat-labile blocking groups on some of the amino acid residues of FastStart Taq DNA Polymerase make the modified enzyme inactive at room temperature. Therefore, there is no elongation during the period when primers can nonspecifically bind. The FastStart Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a preincubation step at 95°C).