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High Fidelity Polymerase

Kód produktu: PCR-204L Kód výrobce: PCR-204L Výrobce: Jena Bioscience GmbH
8 868,57 Kč
7 329,40 Kč bez DPH
do týdne

High Fidelity Polymerase

Zobraz detailní popis

Detailní popis

Thermus species, recombinant, E. coli

For general laboratory use.

Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTP into an acid-insoluble form in 30 minutes at 74 °C.

Shipping: shipped on gel packs

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

Shelf Life: 12 months

Form: liquid

Concentration: 2.5 units/μl

Description:
High Fidelity Pol is based on a blend of Taq DNA polymerase and a proofreading enzyme specially designed for highly accurate and efficient amplification. It shows excellent results with extremely long (up to 30 kb), GC-rich or other difficult templates.
The enzyme blend includes a highly processive 5'→3' DNA polymerase and possesses a 5'→3' polymerization-dependent exonuclease replacement activity. Its inherent 3'→5' exonuclease proofreading activity results in a greatly increased fidelity of DNA synthesis compared to Taq polymerase.
The enzyme is highly purified and free of bacterial DNA.

Fidelity of the enzyme:
High Fidelity Pol is characterized by a 4-fold higher fidelity compared to Taq polymerase.
ERHigh Fidelity Pol = 3.4 x 10-6
The error rate (ER) of a PCR reaction is calculated using the equation ER = MF/(bp x d), where MF is the mutation frequency, bp is the number of base pairs of the fragment and d is the number of doublings
(2d = amount of product / amount of template).

Content:
High Fidelity Pol (red cap)
2.5 units/μl High Fidelity Polymerase in storage buffer

High Fidelity Buffer (green cap)
10x conc.

Recommended 50 μl PCR assay:

5 μl 10x High Fidelity Buffer green cap
200 μM each dNTP -
0.2 - 0.5 μM each Primer -
1 - 100 ng template DNA -
0.5 μl
(1.25 units)
High Fidelity Pol red cap
Fill up to 50 μl PCR-grade water -


Please note that it is essential to add the polymerase as last component.
Recommended cycling conditions:

initial
denaturation
95 °C 2 min 1x
denaturation 95 °C 20 sec 20-30x
annealing1) 50 - 68 °C 30 sec 20-30x
elongation2) 68 °C 1 min/kb 20-30x
final
elongation
68 °C 1 min/kb 1x


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

 

BIOZ Product Citations:
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