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Hot Start Master

Master mix of heat-activatable DNA polymerase for high specificity

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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: 5x conc.

Hot Start Master contains all reagents required for PCR (except template and primer) in a premixed 5x concentrated ready-to-use solution.
The master mix provides improved specificity and sensitivity when amplifying low-copy-number targets in complex backgrounds or when prolonged room-temperature set up is required. The polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during PCR setup.

Activation step:
Hot Start Master requires no prolonged heating or denaturing step. The polymerase inhibiting ligand is quickly released at the increased temperature of thermal cycling.

5x Hot Start Master (red cap)
master mix of heat-activatable DNA polymerase, dATP, dCTP, dGTP, dTTP, KCl, MgCl2 and stabilizers.

PCR grade water (white cap)

Recommended 50 μl PCR assay:

10 μl5x Taq Master Mixred cap
0.2 - 1 μMeach Primer-
2 - 50 ngTemplate DNA-
Fill up to 50 μlPCR grade Waterwhite cap

Recommended cycling conditions:
94 °C2 min1x
Denaturation94 °C30 sec30x
Annealing1)45 - 68 °C30 sec30x
Elongation2)72 °C30 sec - 4 min30x
72 °C2 min1x
1)The annealing temperature depends on the melting temperature of the primers used.
2)The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kbp 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.

Availability Restriction: Exclusively distributed in Japan by Greiner BioOne