Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C

Avoid freeze/thaw cycles.  Hot Start Taq Pol Master Mix (2X) is also stable for three months at 4°C, so for frequent use, an aliquot may be kept at 4°C.

Validade: 12 months

 

2x Hot Start Taq Pol Master Mix Green (purple cap)
Master mix of thermostable Hot Start DNA polymerase, dATP, dCTP, dGTP, dTTP, KCl, MgCl₂ dye, gel loading buffer and stabilizers.

 

Hot Start Taq Pol Master Mix contains Hot Start Taq Polymerase in an optimized PCR buffer with Mg²⁺ and dNTPs. The master mix is supplemented with tracking dyes for direct loading of PCR products on gels. It contains all reagents required for PCR (except template and primer) in a premixed 2x concentrated ready-to-use solution. The Master Mix is recommended for use in routine PCR reactions. It is optimized for high specificity and guarantees minimal by-product formation. Antibody-based hot start technology avoids nonspecific amplification and enables room temperature reaction setup. The tracking dyes in the master mix do not interfere with PCR performance and are compatible with downstream applications such as fluorescent automatic DNA sequencing, ligation, and restriction digestion. The blue dye migrates with 3-5 kb fragments and the yellow dye migrates faster than 20 bp (1% agarose gel).

 

Use the quantities below to prepare a single 50 µl PCR reaction. Thaw, mix, and briefly centrifuge each component before use. Add the following components to a microcentrifuge tube:

 

Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.

Mix and briefly centrifuge the components.

 

Component	50 μl rxn	final conc.
Forward primer (10 μM)	0,5 – 2,5 μl	0,1 – 0,5 μM
Reverse primer (10 μM)	0,5 – 2,5 μl	0,1 – 0,5 μM
Template DNA	x μl	10 pg – 1 μg*

*genomic DNA: 1 ng – 1 μg; plasmidial DNA: 1 pg – 1 ng

 

Cap each tube, mix, and briefly centrifuge the content.

 

Recommended cycling conditions:

**The annealing temperature depends on the melting temperature of the primers used

***The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.

 

For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.