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Multiplex PCR Master

Master mix for multiplex PCR

Product Cat. No. Amount Price (EUR) Buy / Note
Multiplex PCR Master PCR-110S 2 x 1,25 ml (2x conc.) 170,00 Add to Basket/Quote Add to Notepad
Multiplex PCR Master PCR-110L 10 x 1,25 ml (2x conc.) 680,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

Shipping: shipped on blue ice

Storage Conditions: store at -20 °C
avoid freeze/thaw cycles

Shelf Life: 12 months

Form: liquid

Concentration: 2x conc.

Availability Restriction: Exclusively distributed in Japan by Greiner BioOne

Description:
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 routine 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.

Kit contents:
2x Multiplex PCR Master (red cap)
master mix containing Hot Start Taq polymerase, nucleotides, optimized reaction buffer and stabilizers

PCR grade water (white cap)

Recommended 50 μl PCR assay:
Prepare a master mix of all components except template to reduce pipetting errors. A reaction volume of 20-50 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. No-template controls should be included in all amplifications.

componentstock conc.final conc.50 μl
assay
Multiplex PCR Master2x1x25 μl
forward primer 110 μM400 nM2 μl
reverse primer 110 μM400 nM2 μl
forward primer 210 μM400 nM2 μl
reverse primer 210 μM400 nM2 μl
forward primer ...10 μM400 nM2 μl
reverse primer ...10 μM400 nM2 μl
Template
a) animal
genomic DNA
b) bacterial
genomic DNA
c) plasmid and lambda DNA
--
a) 10-200 ng
b) 1 - 50 ng
c) 1 - 5 ng
PCR-grade water--fill up to
50 μl

Recommended cycling conditions:

Initial
denaturation
95 °C12 min1x
Denaturation95 °C30 sec30 - 50x2)
Annealing1)58 - 64 °C40 sec30 - 50x2)
Elongation3)72 °C1 min/kb30 - 50x2)
Final
elongation
72 °C5 min1x
1)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)Cycle numbers are recommended as following:

  • animal genomic DNA
    10 - 50 ng: 35 - 50 cycles
    50 - 200 ng: 30 - 45 cycles
  • bacterial genomic DNA
    1 - 5 ng: 35 - 50 cycles
    5 - 50 ng: 30 - 40 cycles
  • plasmid and lambda DNA
    1 - 5 ng: 30 - 40 cycles

3)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.