A PCR master mix is a prepared mixture that contains all of the components for a PCR reaction that are not sample-specific. You just need to add a DNA template, primers and water.

5x HOT FIREPol® Blend Master Mix Ready to Load is an optimised ready-to-use PCR master mix for PCR assays requiring higher fidelity amplification and hot start functionality. 

It contains a chemically deactivated FIREPol® DNA Polymerase that prevents the possibility of mispriming or primer dimer formation during room temperature setup. The polymerase is reactivated with a 12 – 15 min heating step as part of the initial denaturation step during PCR.

It also contains two carefully optimized enzymes: HOT FIREPol® DNA polymerase and a proofreading polymerase. The enzyme blend has 5’→ 3’ exonuclease activity and 3’→ 5’ proofreading activity for increased copy fidelity. This results in an increased fidelity (up to five fold) compared to regular Taq polymerase. Generated PCR products are compatible with blunt-end and T/A cloning procedures.

This master mix also contains dNTPs, MgCl₂, BSA, and buffer.

• Convenient: ready to load directly onto gels after PCR. No need to add loading dye
• Stable: thermostable polymerase enzyme for room temperature set up, shipping, and short-term storage (up to 1 month if unopened)
• All-in-one format: reduces pipetting errors and saves time
• High fidelity: proofreading activity allows increased copy fidelity
• Hot start functionality: reduced primer dimers and mispriming
• BSA included to help deal with PCR inhibitors present in more difficult DNA templates
• Safe: contains no substances considered hazardous to health at their given concentrations
• Versatile: Suitable for templates up to 5 kb

• Suitable for both routine PCR and more demanding PCR applications
• Ideal for room temperature PCR setup due to hot start modifications
• Ideal for high fidelity applications, such as those involving DNA sequencing, due to proofreading functionality
• Compatible with standard PCR instruments
• Works great as an alternative to other commercial DNA Polymerase master mixes
• Not recommended for applications involving post-PCR spectrophotometric measurements (e.g. Sanger sequencing) without downstream dye cleanup, as loading dyes may interfere with measurements

Recommended usage

We recommend using 4 μL of 5x PCR master mix per 20 μL reaction.

Components

• HOT FIREPol® DNA polymerase: chemically modified FIREPol® DNA Polymerase enabling hot start
• Proofreading enzyme: to enhance copy fidelity – up to 5x that of standard Taq polymerases
• 5x Blend Master Mix Buffer with 7.5 mM MgCl2: 1x PCR solution contains 1.5 mM MgCl2
• 1 mM dNTPs of each: 1x PCR solution contains 200 µM dATP, 200 µM dCTP, 200 µM dGTP and 200 µM dTTP
• BSA
• Blue dye: migration equivalent to 3.5 – 4.5 kb DNA fragment
• Yellow dye: migration equivalent to 35 – 45 bp DNA fragment
• Compound that increases sample density for direct loading

Storage & Stability

Routine storage at –20 ºC.

Temporary storage for up to 1 month at room temperature, or up to six months at 2 – 8 ºC, has no detrimental effects on the quality of HOT FIREPol® Master Mix Ready to Load.

Shipping conditions

Shipped at room temperature.