Reagent	Volume	Comment
5x PCR buffer	1,450 µl
PCR additive	1,450 µl
2 mM dNTPs	725 µl	2 mM each nucleotide
50 µM AatII_F or AvrII_R primer	145 µl
1 ng/µl pBR322_AvrII plasmid	145 µl
2 units/µl DNA polymerase	72.5 µl
water	363 µl