Whole plasmid amplification by PCR
Posted 23 July 2013 - 06:48 AM
is it possible to amplify an entire plasmid by PCR so as to get enough material for mammalian cell transfection?
My specific problem is that my plasmid has a highly repetitive insert several kilobases in lenght so conventional amplification by cloning in bacteria is unpractical.
Is there any highly processive Taq-pol with proof-reading activity that I might use for this amplification?
Thanks in advance for your answers!
Posted 23 July 2013 - 12:37 PM
KOD and pfu polymerases are ones commonly used for this sort of thing.
Posted 02 August 2013 - 01:16 AM
i have used Pfu polymerase to amplify whole plasmid for SDM.. works fine