HOT FIREPol® MultiPlex Mix Ready To Load with 10 mM MgCl2

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

Generated PCR products are compatible with T/A cloning procedures.

Recommendations

We recommend using HOT FIREPol^® MultiPlex Mix Ready To Load in any PCR application that will be visualized by agarose gel electrophoresis and ethidium bromide staining.

HOT FIREPol^® MultiPlex Mix Ready To Load is not recommended for use in applications where spectrophotometric 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: 1 x Taq

Cloning Type: T/A cloning 

Multiplexing: tested with up to 18 targets (depending on amplicon length)

Amplicon Size: up to 5 kb

Mix Components

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

5x Multiplex Buffer with 10 mM MgCl₂: 1x PCR solution – 2 mM MgCl₂

dNTPs: dATP, dCTP, dGTP and dTTP

Bovine serum albumin (BSA) to enhance PCR reaction

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