Team:Freiburg/Protocols/PCR Digest
PCR Template Digest
protocol for digestion of the PCR template with DpnI
DpnI needs a methylation at its recognition site (GATC) in order to recognize it. Therefore only the template, which was amplified in E. coli and thus is methylated will be digested, while the unmethylated PCR products will remain intact.
material: DpnI, thermoblock, PCR product
duration: 10 min preparation, 80 min incubation
- Add 2 µl DpnI to your PCR products
- Incubate at 37°C for 60 min (digest)
- Incubate at 80°C for 20 min (heat inactivation of DpnI)
- PCR products can be cleaned-up and used e.g. for cloning purposes
