"
Team:Newcastle/Restriction digests
From 2010.igem.org
(Difference between revisions)
(→Restriction digestion) |
|||
| Line 2: | Line 2: | ||
=Restriction digestion= | =Restriction digestion= | ||
| - | |||
| - | |||
==Materials required== | ==Materials required== | ||
Revision as of 08:50, 30 July 2010
| |||||||||||||
| |||||||||||||
Contents |
Restriction digestion
Materials required
- Eppendorf tubes
- Pipettes
- Appropriate DNA/plasmid
- Appropriate restriction enzymes
- 10X buffer
- Water
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 should be used for a single reaction - glycerol inhibits reaction
- 10x buffer must be diluted to 1x i.e. 10% final volume
   
|
    
|
