Restriction digestion

To cut DNA at specific base sequences using restriction enzymes

Procedures

1. Make up 20 µl reaction mix as follows:
   • 15 µl of DNA/plasmid
   • 1 µl of restriction enzyme 1
   • 1 µl of restriction enzyme 2
   • 2 µl of 10X buffer
   • 1 µl of water
2. Incubate at 37°C for 3 hours

Notes

• No more than 10% of enzyme should be used for a single reaction - glycerol inhibits reaction
• 10x buffer must be diluted to 1x i.e. 10% final volume