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Team:Newcastle/Restriction digests
From 2010.igem.org
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==Procedures== | ==Procedures== | ||
| - | # Make up | + | # Make up 20 µl reaction mix as follows: |
| - | *1 µl of restriction | + | *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 | ||
# Incubate at 37°C for 3 hours | # Incubate at 37°C for 3 hours | ||
Revision as of 15:14, 28 July 2010
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Restriction digestion
To cut DNA at specific base sequences using restriction enzymes
Procedures
- 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
- Incubate at 37°C for 3 hours
Notes
- No more than 10% of enzyme - solution contains glycerol which inhibits reaction
- 10x buffer must be diluted to 1x i.e. 10% final volume
   
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