KAPA Long Range PCR Kits
Amplify long templates up to 20 kb with extreme sensitivity and increased fidelity.
The KAPA Long Range PCR system is a blend of Taq DNA polymerase and an engineered archaeal (B-family) DNA polymerase possessing proofreading capability. This two-enzyme system is designed specifically to support long range and/or sensitive PCR.
KAPA Long Range DNA Polymerase possesses 3x – 4x higher fidelity than Taq polymerase. The majority of PCR products from a KAPA Long Range HotStart DNA Polymerase reaction have a single deoxyadenosine added to the 3’ ends of the amplicon and can be used for ligation into single 3’-T -overhang cloning sites.
The KAPA Long Range PCR system is ideally suited for:
- PCR amplification of long targets up to 20 kb and/or PCR using low amounts 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.
- Targeted sample enrichment for next-generation DNA sequencing.
KAPA Long Range DNA Polymerase is also available with HotStart technology for improved specificity during prolonged benchtop setup or liquid handling.
Kapa Biosystems enzymes outperform others we've used. In particular, KAPA Long Range HotStart DNA Polymerase has enabled us to reliably amplify long range templates up to 12 kb for use in downstream next-generation DNA resequencing. KAPA Long Range HotStart outperformed three other competitors' enzymes in our head-to-head comparisons. Kapa enzymes are the cornerstone of our current work and we use KAPA Long Range HotStart, KAPA2G Fast HotStart, and KAPA HiFi because of their exceptional performance and great value.
- Keith Choate, M.D., Ph.D., Department of Dermatology, Yale University School of Medicine, New Haven, Connecticut
The KAPA Long Range 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. KAPA Long Range possesses 3x - 4x higher fidelity than Taq polymerase. KAPA HiFi DNA Polymerase is recommended for short and mid-range targets where higher fidelity is required.
All KAPA Long Range kits are supplied with 5x KAPA Long Range 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.
The KAPA Long Range 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
- Targeted resequencing using next-generation DNA sequencing
Long Range and High Sensitivity
KAPA Long Range 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.
KAPA Long Range 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.
KAPA Long Range 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.