KAPALongRange

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The KAPALongRange PCR system is a blend of Taq DNA polymerase and a modified archaeal (Type B) DNA polymerase possessing proofreading capability. This two-enzyme system is designed specifically to support long range and/or sensitive PCR. The KAPALongRange system polymerizes DNA from a primer annealed to a DNA template in the presence of deoxyribonucleotide triphosphates. Both enzymes possess 5’→3’ polymerase activity, but only Taq possesses double strand dependent 5’→3’ exonuclease activity and only the Type B polymerase possesses 3’→5’ exonuclease (proofreading) activity.

KAPALongRange possesses higher fidelity than Taq polymerase. KAPAFidelity is recommended for shorter targets where higher fidelity is required.

All KAPALongRange kits are supplied with 5x KAPALongRange Buffer and 250 µl KAPA dNTPs (10 mM each). This buffer has been optimized for demanding applications that require long range and high sensitivity. Magnesium chloride solution is supplied separately to accommodate PCR optimization.

Product Applications

The KAPALongRange PCR system is suited for:

  • PCR amplification of long targets and/or PCR using low concentrations of template DNA
  • Standard short- and mid-range PCR amplification
  • Production of PCR products to be used for ligation into 3’-T-overhang cloning vectors

LONG RANGE AND HIGH SENSITIVITY

KAPALongRange is able to amplify long templates from limiting complex genomic DNA.

Figure 1 Amplification of 15 kb fragment (tPA gene) from human chromosomal DNA. Lanes: (M) Marker, (1) 25 ng human gDNA, (2) 5 ng human gDNA, (3) 1 ng human gDNA. 35 cycles, 68ºC extension temperature, 2.5 units of enzyme.

KAPALongRange exhibits high yield, greater sensitivity and specificity compared with Roche Expand™ Long Template and Takara LA Taq™.

Figure 2 Amplification of 15 kb fragment from 10 pg, 10 fg, and 1 fg of lamdba DNA. Lanes: (1-3) KAPALongRange, (M) Marker, (4-6) Roche Expand, (7-9) Takara LA.

KAPALongRange can replace Taq polymerase using standard PCR cycling conditions and less enzyme per reaction. KAPALongRange amplifies mid-range and long-range targets at greater sensitivity and specificity as compared with Taq polymerase and Takara Ex Taq™.

Figure 3 Amplification of 6 log dilution series of lambda DNA starting with 1 ng to 10 fg. Amplicons were 2 kb, 8 kb, and 15 kb in length. 35 cycles, 72ºC extension temperature, 1.25 units of enzyme.

Figure 4 Amplification of 15 kb fragment from 10 pg, 10 fg, and 1 fg of lamdba DNA. Lanes: (1-2) KAPALongRange, (M) Marker, (3-4) Takara Ex Taq™. 35 cycles, 68ºC extension temperature, 1.25 units of enzyme.

Kit Components:

  • 5 U/µl KAPALongRange Polymerase
  • 5x KAPALongRange Buffer
  • 25 mM MgCl2 solution
  • 250 µl KAPA dNTP Mix (10 mM each)
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