Specialty PCR Enzymes

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D7442 MTP Taq DNA Polymerase Taq DNA Polymerase, free of DNA contaminants MTP Taq DNA Polymerase is a recombinant thermostable enzyme from Thermus aquaticus expressed in E. coli and purified using a proprietary process to minimize levels of contaminating DNA. The enzyme has both 5′→3′ DNA polymerase and exonuclease activities, is ∼95 kDa by SDS-PAGE, and has no detectable endonuclease or 3′→5′ exonuclease activities. Each lot of MTP Taq undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA.

Contaminating DNA present in most other polymerase preparations often preclude or obscure the accurate interpretation of results, especially when targeting conserved sequences, e.g., bacterial 16S rRNA region. Through Sigma′s proprietary DNA removal methods and strict quality control standards, we can ensure the absence of the most commonly found contaminant DNA. Each lot of MTP Taq is assayed using PCR and primers specific to (1) the conserved region of bacterial 16S rRNA, (2) the Taq expression vector, and (3) the human β-actin gene.

While MTP Taq ensures a high-quality, low contaminant DNA polymerase for reliable PCR amplification, DNA contaminants can be introduced into PCR through a number of other reagents. To further minimize the risk of contaminant DNA during PCR, we include 10× MTP Taq Buffer with each tube of MTP Taq DNA Polymerase. Each lot of 10× MTP Taq Buffer undergoes the same strict quality control testing as MTP Taq DNA polymerase to ensure the absence of contaminating DNA. To prevent false positive PCR results, only DNA-free reagents should be used in PCR reactions with MTP Taq DNA polymerase.
R1028 Restorase® DNA Polymerase with 10× Reaction Buffer Enzyme blend for PCR amplification of damaged DNA Restorase DNA Polymerase with 10× Reaction Buffer combines Sigma′s long and accurate enzyme technology with a small amount of DNA repair enzyme. The optimized blend will initiate the repair and further amplification of damaged DNA templates greater than 800 bp. Restorase has also been shown to increase yield on undamaged DNA templates.
GE25-6400-10 TempliPhi 100 Amplification Kit GE Healthcare, 25-6400-10 illustra TempliPhi DNA Amplification Kits use isothermal rolling circle amplification (RCA) to efficiently prepare DNA sequencing templates.

TempliPhi Kits use a unique process to efficiently prepare micrograms of circular DNA from picogram input material. The DNA templates are prepared by rolling circle amplification (RCA) using bacteriophage Phi29 DNA polymerase. TempliPhi uses an isothermal method for the exponential amplification of circular DNA. Phi29 DNA polymerase is active at 30° C, enabling amplification to be performed at this temperature without the need for thermal cycling. The TempliPhi protocol requires less than 20 min of hands-on time to amplify 96 samples from bacterial colonies.

Two different types of TempliPhi DNA Amplification Kits are available to meet different throughput requirements. illustra TempliPhi DNA Amplification Kit for 2000 reactions contains premixed components and generates templates after overnight incubation. The 100- and 500-reaction kits contain separate, unmixed components for enhanced stability.

The illustra TempliPhi DNA Sequencing Template Amplification Kit has a distinct protocol and workflow compared to the 100/500 reaction size. Procedures for each kit are not interchangeable. Prior optimization is needed.
GE25-6400-50 TempliPhi 500 Amplification Kit GE Healthcare, 25-6400-50 illustra TempliPhi DNA Amplification Kits use isothermal rolling circle amplification (RCA) to efficiently prepare DNA sequencing templates.

TempliPhi Kits use a unique process to efficiently prepare micrograms of circular DNA from picogram input material. The DNA templates are prepared by rolling circle amplification (RCA) using bacteriophage Phi29 DNA polymerase. TempliPhi uses an isothermal method for the exponential amplification of circular DNA. Phi29 DNA polymerase is active at 30° C, enabling amplification to be performed at this temperature without the need for thermal cycling. The TempliPhi protocol requires less than 20 min of hands-on time to amplify 96 samples from bacterial colonies.

Two different types of TempliPhi DNA Amplification Kits are available to meet different throughput requirements. illustra TempliPhi DNA Amplification Kit for 2000 reactions contains premixed components and generates templates after overnight incubation. The 100- and 500-reaction kits contain separate, unmixed components for enhanced stability.

The illustra TempliPhi DNA Sequencing Template Amplification Kit has a distinct protocol and workflow compared to the 100/500 reaction size. Procedures for each kit are not interchangeable. Prior optimization is needed.