How do I DpnI digest my PCR reaction?
Add 1-2 μl directly to the PCR reaction (works well in PCR buffer) after it’s cooled down, and incubate for 1 hour at 37°C, followed by gel-purification. Use as little template as possible for the PCR reaction – 1-2 ng plasmid, and 30-35 cycles. This way the risk of plasmid contamination is minimal. If you use the same template over and over again, you could use a little bit of purified PCR-band as template instead – this way you’ll have no problem with plasmid contamination. You should still gel-purify, however, to avoid other contaminations.
