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Hot Start Core Kit

Kit of heat-activatable DNA polymerase for high specificity, dNTPs and reaction buffer, aptamer-blocked

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For in vitro use only!

Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dNTP's into an acid-insoluble form in 30 minutes at 70 °C using hering sperm DNA as substrate.

Shipping: shipped on blue ice

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

Shelf Life: 12 months

Form: liquid

Concentration: 5 units/μl

Hot Start Core Kit contains all reagents required for PCR (except template and primer) in one box combining simple handling with high flexibility. The remium quality polymerase, ultrapure dNTPs and the optimized complete reaction buffer ensure superior amplification results. The additional reaction buffer without MgCl2 in combination with the MgCl2 stock solution allows an easy optimization of difficult amplifications. The kit 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-dimer formation at low temperatures during PCR setup. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in 5'→3' direction in the presence of magnesium. It also possesses a 5'→3' polymerization-dependent exonuclease replacement activity but lacks a 3'→5' exonuclease activity.

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


Hot Start Pol*
5 units/μl
red cap
40 μl
200 units
200 μl
1000 units
dNTP Mix
10 mM each
white cap
200 μl1 ml
Hot start Buffer complete**
10x conc.
green cap
1.2 ml5 x 1.2 ml
MgCl2 Stock
25 mM
yellow cap
1.5 ml2 x 1.5 ml

*in 20 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5 % Tween-20, 0.5 % Nonidet P-40, 50 % (v/v) Glycerol, pH 8.0 (25°C)
**200 mM Tris-HCl, 500 mM KCl, 15 mM MgCl2, pH 8.5 (25°C)

Recommended 50 μl PCR assay:

5 μl10x Hot Start Buffer completegreen cap
1 μldNTP Mixwhite cap
1-5 μleach Primer
(10 μM)
2 - 50 ngtemplate DNA-
0.2 - 0.5 μl
(1 - 2.5 units)
Hot Start Polred cap
Fill up to 50 μlPCR-grade water-

Optimization of MgCl2 concentration:
A concentration of 1.5 mM Mg2+ is recommended for most applications. For an individual optimization add MgCl2 stock solution as shown in the table below.

50 μl PCR assay

MgCl2 Stock Solution1 μl2 μl3 μl5 μl
final MgCl2 conc.2 mM2.5 mM3 mM4 mM

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/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.

Related products:

  • Ready-to-Use Mixes / direct gel loading
  • Ready-to-Use Mixes
  • Thermophilic Polymerases
  • Deoxynucleotides (dNTPs)
  • Supplements
  • Primers and Oligonucleotides
  • DNA Ladders

Availability Restriction: Exclusively distributed in Japan by Greiner BioOne