Team:Newcastle/Restriction digests
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==Notes== | ==Notes== | ||
*No more than 10% of enzyme should be used for a single reaction - glycerol inhibits reaction | *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 | + | *10x buffer must be diluted to 1x i.e. 10% final volume. |
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+ | '''Go back to our [[Team:Newcastle/Protocol list|Protocol List]]''' | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} |
Revision as of 09:15, 30 July 2010
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Contents |
Restriction digestion
Materials required
- Eppendorf tubes
- Pipettes
- Appropriate DNA/plasmid
- Appropriate restriction enzymes
- 10X buffer
- Water
Procedures
Add the following as mentioned below to make up to a final volume of 20 µl of reaction mix:
- 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 the digestion mix 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.
Go back to our Protocol List