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Team:Newcastle/Restriction digests
From 2010.igem.org
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{{Team:Newcastle/mainbanner}} | {{Team:Newcastle/mainbanner}} | ||
| + | =Restriction digestion= | ||
To cut DNA at specific base sequences using restriction enzymes | To cut DNA at specific base sequences using restriction enzymes | ||
| - | == | + | ==Procedures== |
| - | + | # Make up 30 µl reaction mix as according: | |
*1 µl of restriction enzymes in total | *1 µl of restriction enzymes in total | ||
*3 µl of 10x buffer | *3 µl of 10x buffer | ||
*remainder is DNA/water | *remainder is DNA/water | ||
| + | # Incubate at 37°C for 3 hours | ||
| - | + | ==Notes== | |
*No more than 10% of enzyme - solution contains glycerol which inhibits reaction | *No more than 10% of enzyme - solution contains glycerol which inhibits reaction | ||
*10x buffer must be diluted to 1x i.e. 10% final volume | *10x buffer must be diluted to 1x i.e. 10% final volume | ||
| - | + | {{Team:Newcastle/footer}} | |
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Revision as of 15:12, 28 July 2010
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Restriction digestion
To cut DNA at specific base sequences using restriction enzymes
Procedures
- Make up 30 µl reaction mix as according:
- 1 µl of restriction enzymes in total
- 3 µl of 10x buffer
- remainder is DNA/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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