Long-range PCR with a DNA polymerase fusion.
Proofreading DNA polymerase fusions offer several advantages for long-range PCR, including faster run times and higher fidelity compared with Taq-based enzymes. However, their use so far has been limited to amplification of small to mid-range targets. In this article, we present a modified protocol for using a DNA polymerase fusion to amplify genomic targets exceeding 20 kb in length. This procedure overcomes several limitations of Taq blends, which up until recently, were the only option for long-range PCR. With a proofreading DNA polymerase fusion, high-molecular-weight amplicon can be generated and analyzed in a single day, and a significant proportion is expected to be error-free.
Hogrefe
Holly H
HH
Stratagene Products Division, Agilent Technologies, Inc, La Jolla, CA, USA.
Borns
Michael C
MC
D001483:Base Sequence; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase