Description

5x HOT FIREPol^® Blend Master Mix Ready to Load is a premixed ready-to-use solution containing all reagents required for PCR (except template, primers and water), additional compound needed for direct loading onto agarose gel and two tracking dyes (blue and yellow) that allow to monitor progress during electrophoresis.

HOT FIREPol^® Blend Master Mix contains two carefully optimized enzymes – HOT FIREPol^® DNA polymerase and a proofreading polymerase. This enzyme blend has both the 5’→ 3’ exonuclease activity as well as the 3’→ 5’ proofreading activity. HOT FIREPol^® Blend Master Mix exhibits an increased fidelity (up to five fold) compared to regular Taq polymerase.

In addition, the mix also allows amplification of longer fragments.

Generated PCR products are compatible with blunt-end and T/A cloning procedures.

HOT FIREPol^® Blend Master Mix exhibits an increased fidelity (up to five fold) compared to regular Taq polymerase.

Recommendations

We recommend using 5x HOT FIREPol^® Blend Master Mix Ready to Load in any PCR application that will be visualized by agarose gel electrophoresis and ethidium bromide staining.

5x HOT FIREPol^® Blend Master Mix Ready to Load is not recommended for use in applications where spectro-photometric measurements (absorbance or fluorescence) are necessary because yellow and blue dyes can interfere with these applications.

Properties

Concentration: 5x

Hot-start: yes, initial activation in 12-15 min

Ready to load: yes

Fidelity: 5 x Taq

Cloning Type: T/A cloning and Blunt-end cloning

Amplicon Size: up to 5 kb

Mix Components

HOT FIREPol^® DNA polymerase: chemically modified FIREPol^® DNA Polymerase enabeling hot-start

Proofreading enzyme: to enhance fidelity

5x Blend Master Mix Buffer with 7. 5 mM MgCl₂: 1x PCR solution – 1.5 mM MgCl₂

1 mM dNTPs of each: 1x PCR solution – 200 µM dATP, 200 µM dCTP, 200 µM dGTP and 200 µM dTTP

Bovine serum albumine (BSA) to enhance PCR reaction

Blue dye: migration equivalent to 3.5-4.5 kb DNA fragment

Yellow dye: migration rate in excess of primers in 1% agarose gel: <35-45 bp

Compound that increases sample density for direct loading