Which polymerase is good for amplifying 4.5kb fragment from a plasmid?
There are lots of great polymerases available that can handle this length of an amplification
I like Q5 from New England Biolabs, it's proofreading, can do very long extensions, and leaves blunt ends
Vent is another good option
It depends on what vendors are available, many of them will also give you a small test sample for free so you can try out their products before buying
You could also try Pfu Turbo from Agilent, which is commonly used to propagate plasmids from site-directed mutagenesis reactions; I used this to generate PCR products anywhere from ~1 kb to up to ~5-5.5 kb in length.
As the previous scientist told, there are many good polimerases able to work well in this size range. I reccoment you an high fidelity since the size of the fragment is high, with a low fidelity you have high probability to introduce some mutations.
i like a lot:
-kapa hifi (Roche)
- CLone amp (Promega)
that i'm routinelly using to perform vector PCR in pipe cloning for gene coloning or mutagenesis.
I the past i used also pfu ultra fusion II and also this work well
Q5 in my experience work better for smaller fragments but i did not work a lot with it.
best regards
Manuele
Hi,
In my lab we are usually using Q5 or Phusion high-fidelity polymerases for amplification of longer fragments.
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