Long-range PCR refers to the amplification of DNA targets from 5 kb – 20 kb in length. Typically, a blend of thermostable DNA polymerases, consisting primarily of Taq polymerase combined with a small amount of proofreading polymerase, is required for efficient amplification of DNA targets >5 kb. Taq polymerase lacks proofreading activity and is unable to efficiently extend beyond misincorporated bases. Mismatched base pairing generates truncated products that accumulate during PCR and contribute to reaction failure if the target is long and/or the template DNA is limiting. The addition of a proofreading polymerase to Taq allows mismatches incorporated at the 3′ end of the growing strand to be repaired. The resulting polymerase blend is, therefore, able to support PCR of longer targets than either enzyme alone.
KAPA Long Range PCR Kits contain a blend of Taq DNA polymerase and a proofreading DNA polymerase. This two-enzyme system is designed specifically to support long-range and sensitive PCR, possessing 3 – 4X higher fidelity than Taq polymerase.
KAPA HiFi DNA Polymerase is recommended for long-range PCR applications (<10 kb) that require high fidelity. KAPA HiFi is an evolved, proofreading polymerase with fidelity 100X higher than Taq polymerase. The KAPA HiFi enzyme is optimized for amplifying longer targets, with higher sensitivity, than other single-enzyme proofreading polymerases.
For Research Use Only. Not for use in diagnostic procedures.