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
