HOT START DNA POLYMERASE

  • PCR Enzymes
  • Taq
  • Glycerol Free Products
  • Hot Start
  • High Fidelity
  • Genotyping
  • Multiplex
  • GC-rich DNA Amplification
  • Real-Time PCR
  • HRM - High Resolution Melting
  • Nucleotides
  • PCR Buffers
  • DNA Loading Buffer
  • PCR Ladders
  • DNA and RNA Extraction
  • PCR Clean-Up

Hot Start

TEMPase Hot Start DNA Polymerase is a modified form of Ampliqon Taq DNA Polymerase activated by heat treatment. A chemical moiety is attached to the enzyme, which makes the enzyme inactive at room temperature.

During setup and the first ramp of cycling the enzyme is not active and misprimed primers are not extended. Once the PCR reaction reaches the optimal activation temperature, the chemical moiety is cleaved releasing the active TEMPase Hot Start DNA Polymerase. Resulting in higher specificity, increased sensitivity and greater yield compared to standard DNA polymerases.

TEMPase Hot Start DNA polymerase is manufactured in-house in accordance to our ISO 9001:2015 quality management system, ensuring that each batch possess the same robust performance every time.

TEMPase Hot Start DNA polymerase is available in different standalone variants and master mixes.

Standalone variants are supplied either without buffers or in kits including MgCl2 and one or two Ampliqon PCR buffers to achieve highest PCR performance and to avoid tedious reaction optimisation.

Chemical inactivation of TEMPase Hot Start DNA Polymerase has proven highly effective compared to other inactivation methods such as antibody inactivation. The chemically modified enzyme withstands longer periods of time at room temperature without non-specific PCR amplifications. This feature is useful if you need pre-incubation at elevated temperatures, for example in case of UNG treatment at 50 °C prior to PCR.

 

TEMPase Hot Start DNA Polymerase

TEMPase Hot Start DNA Polymerase 5 U/µl has been designed to diminish the formation of non-specific priming events during reaction set-up and the first ramp of thermal cycling. TEMPase Hot Start DNA Polymerase therefore enables detection of low abundance targets as well as multiplexing purposes. 

FEATURES

  • Reaction set-up at room temperature
  • dUTP incorporation possible
  • Increased sensitivity, specificity and product yield

DESCRIPTION

This is a chemically modified version of Ampliqon Taq DNA Polymerase and is activated by heat treatment.

A chemical moiety is attached to the enzyme at the active site, which renders the TEMPase Hot Start enzyme inactive at room temperature.
During set up and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended. Once the PCR reaction reaches the optimal activation temperature, the chemical moiety is cleaved releasing the active TEMPase Hot Start DNA Polymerase. Resulting in higher specificity, sensitivity and yield compared to standard Ampliqon Taq DNA polymerase.

This DNA Polymerase is suitable for detection of low abundance targets, screening, multiplexing, direct colony PCR and Real time PCR applications.

TEMPase Hot Start DNA polymerase also exits in glycerol format for automation and lyophilization: TEMPase Hot Start DNA Polymerase Glyceerol Free

TEMPASE HOT START DNA POLYMERASE PROMOTES INCREASED SPECIFICITY AND YIELD

Example of PCR amplification of BAIP3. Taq or TEMPase were used as indicated with Ammonium Buffer at the indicated Mg2+ concentrations. Taq results in specific and high yield band at only one Mg2+ concentration (2 mM). TEMPase results in specific bands over a broad range of Mg2+ concentrations and increased yield. M: Marker.
 

TEMPASE HOT START DNA POLYMERASE IS INACTIVE AT AMBIENT TEMPERATURE

 


Ampliqon TEMPase is activated by initial heating at 95 °C for 15 minutes. (lane 1). Without activation the enzyme is completely inactive (lane 2). M: marker.

PRODUCT INFO

TEMPase Hot Start DNA Polymerase

With Ammonium Buffer, 5 U/µl
With Combination Buffer, 5 U/µl
5x PCR Buffer RED, 5 U/µl
Without buffer, 5 U/µl
Sample, 5 U/µl

TEMPase Hot Start Master Mix

TEMPase Hot Start Master Mix is a ready to use mix that offers easy reaction assembly at room temperature prior to hot start PCR. Just add your template and primers to successfully carry out PCR.
TEMPase Hot Start Master Mix is the popular alternative to TEMPase Hot Start DNA Polymerase. It offers the same excellent performance and increased reproducibility.


FEATURES

  • Minimal optimisation
  • Convenient reaction set up at room temperature
  • Time-saving premixed solution
  • Increased sensitivity, specificity and product yield
  • Consistent test results
  • Lower risk of contamination

DESCRIPTION

Ampliqon TEMPase Hot Start DNA Polymerase 2x Master Mix is a ready to use master mix composed of TEMPase Hot Start DNA Polymerase, dNTPs, MgCland either TEMPase Buffer C (a balanced KCI/(NH4)2 SO4 Tris buffer system) or TEMPase Buffer A (a (NH4)2 SO4 tris buffer system).

The TEMPase buffer system covers all needs for PCR applications and allow the user to choose the optimal buffer for specific PCR set-ups and applications.

TIP – CHOOSE THE RIGHT TEMPASE HOT START MASTER MIX

For most standard applications Master Mix A based on Ammonium Buffer is the best option. It promotes robust amplification, high yield and high specificity. In certain cases, Master Mix C is the best option.

If you want to visualise on agarose gels, we suggest that you choose Master Mix BLUE A or master Mix BLUE C.

COMPARISON OF TEMPASE HOT START POLYMERASE 2X MASTER MIX WITH COMPETITOR

TEMPase Hot Start DNA Polymerase 2x Master Mix A was compared with an equivalent Hot Start master mix from a well-recognised competitor. Four different DNA targets varying in length PAH (203 bp), ENG9 (293 bp), Q8 (727 bp) and ENG5 (480 bp) were evaluated. TEMPase Hot Start DNA Polymerase 2x Master Mix A performed equally well or better than competitor. Each Amplification has been conducted according to supplier’s manual.

PRODUCT INFO

TEMPase Hot Start Master Mix

Master Mix A
Sample
Master Mix C
Sample

TEMPase Hot Start Master Mix BLUE

Taq OptiMix CLEAR

TEMPase Hot Start 2x Master Mix BLUE is a time saving alternative to TEMPase Hot Start 2x Master Mix. It offers the same excellent performance. In addition, the blue loading dye facilitates direct gel loading. You do not need a separate loading buffer and time-consuming sample preparation before electrophoresis. This makes TEMPase Hot Start Master Mix BLUE especially suitable for high throughput testing.


FEATURES

  • Minimal optimisation
  • Time-saving premixed solution
  • Direct loading onto agarose and SDS DNA gels
  • Easy visualisation of pipetting
  • Dye front running at 100-500 bp (0.5 -1.5 % agarose gel)
  • Convenient reaction set-up at room temperature
  • Increased sensitivity, specificity and product yield
  • Consistent test results
  • Lower risk of contamination

DESCRIPTION

This is a ready-to-use 2x master mix composed of dNTPs, a blue loading dye, a stabiliser, MgCl2 and either TEMPase Buffer C (a balanced KCl / (NH4)2 SO4 buffer system)  or TEMPase Buffer A (a (NH4)2 SO4 buffer system).

The blue dye and stabiliser do not interfere with the PCR. If necessary, the blue dye can be removed by spin column purification or other methods. Furthermore, PCR products generated using TEMPase Hot Start DNA Master Mix BLUE, can directly be used for sequencing after treatment with either spin columns or PureIT ExoZAP PCR CleanUp. 

 

TIP – CHOOSE THE RIGHT TEMPASE HOT START MASTER MIX

For most standard applications Master Mix BLUE A based on Ammonium Buffer is the best option. It promotes robust amplification, high yield and high specificity. In certain cases Master Mix BLUE C is the best option.

DIRECT GEL LOADING

After end of amplification, using TEMPase DNA Polymerase Hot Start Master Mix BLUE PCR products can be loaded directly onto agarose and SDS DNA gels. The blue dye provides easy visualisation of both pipetting and gel loading. The blue dye front runs at 100 – 500 bp on 0.5 – 1.5 % agarose gels.

PRODUCT INFO

TEMPase Hot Start Master Mix BLUE

Master Mix A
Sample
Master Mix C
Sample