1、PCR reaction system:
     25 ng linear template (~6.5 kb)
     50 pmol each primer
     100 pmol each dNTP
     1X Promega Taq buffer (no Mg2+)
     1.5 mM MgCl2
     1 U Taq DNA polymerase in 50 ul final volume

2、PCR programme:
    92°C / 2'
    92°C / 30"
    50°C or 55°C (depends on Tm of oligos) /30"
    72°C / about 2' per kb
    Go to 2, 15 times
    70°C/ 8' 
    4°C,hold.
    Takes about 2 hours to complete.

3、Notes:

If you are using Pfu turbo, use its buffer (Mg already added) and decrease elongation to 1'/kb DNA. Note that this DNA is blunt ended and can be cloned directly (no purification necessary) into a phosphorylated vector. Taq made DNA needs to be AT vector cloned.