DNA polymerases for long fragment amplification (Long Range PCR)
The polymerase chain reaction (PCR) has been used extensively in the
amplification of DNA fragments where there is a general size preference
for products less than 1000 bp. Larger products may be synthesized at
low or inefficient levels and then normally after considerable effort
in reaction optimization. The maximum amplifiable length of PCR is
limited by the low fidelity of the Thermus aquaticus (Taq) DNA
polymerase, the most commonly used thermostable polymerase. This
limitation is thought to be due to the error‐prone nature of the Taq
polymerase, whereby those transcripts that end in an error (a
misincorporated base) are not efficiently reinitiated and extended.
Long range PCR allows the amplification of PCR products, which are much larger than those achieved with conventional Taq polymerases. Up to 27 kb fragments are possible from good quality genomic DNA. Amplification of long DNA fragments is desirable for numerous applications such as physical mapping applications and direct cloning from genomes.
Long range PCR allows the amplification of PCR products, which are much larger than those achieved with conventional Taq polymerases. Up to 27 kb fragments are possible from good quality genomic DNA. Amplification of long DNA fragments is desirable for numerous applications such as physical mapping applications and direct cloning from genomes.