Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C

Avoid freeze/thaw cycles.

Validade: 12 months

 

2x Multiplex PCR Master (purple cap)
Master mix of thermostable Hot Start DNA polymerase, dATP, dCTP, dGTP, dTTP, KCl, MgCl₂  and stabilizers.

 

 

Multiplex PCR Master is specially designed for the set-up of multiplex PCR reactions. It contains an optimized composition of polymerase, nucleotides, MgCl₂ and stabilizing components in a specifically developed buffer system allowing the parallel amplification of a multitude of fragments in a single PCR assay. The master mix contains all reagents (except primer and template) in a 2x concentrated ready-to-use solution. The kit is recommended for use in clinical PCR reactions and highly suitable for multiple target gene amplification in a single tube. The high specificity and sensitivity of the mix is achieved by a chemically inhibited hot-start polymerase. Its activity is blocked at ambient temperature preventing the extension of nonspecifically annealed primers and primer-dimer formations at low temperatures during PCR setup.

 

A reaction volume of 10-50 µl per assay is recommended for most PCR cyclers. Pipet with sterile filter tips and perform the setup in an area separate from DNA preparation or analysis. Notemplate controls should be included in all amplifications. 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 optimal annealing temperature (AT) can be calculated for each primer as following: AT = Tm - 5 °C with Tm = 2 °C x (A+T) + 4 °C x (G+C) Please note that primers should be designed to show minimal differences in there melting temperatures (Tm). 2)The elongation time d

***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.