Team:Groningen/protocols/Digestion
From 2013.igem.org
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Latest revision as of 23:32, 4 October 2013
Restriction digestion
Fast digest
Materials:
- MQ water
- 1.5 ml tubes
- Fast digestion buffer
- Restriction enzymes
- DNA
Reaction:
Compound | 20 µl | Final concentration |
---|---|---|
MQ water | up to 20 µl | |
10x fast digestion buffer | 2 µl | 1x |
restriction enzyme(s) | 1 µl | 1U |
DNA | 4 µl | up to 1 µg |
When the reaction mix is ready, the content is mixed and incubated at 37°C for 1h [1].
Normal digest
Materials:
- MQ water
- 1.5 ml tubes
- Enzyme digestion buffer
- Restriction enzymes
- DNA
Reaction:
Compound | 20 µl | Final concentration |
---|---|---|
MQ water | up to 20 µl | |
10x restriction digestion buffer | 2 µl | 1x |
restriction enzyme(s) | 0.5 µl | 1U |
DNA | 2 µl | 0.2-1.5 µg |
When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h.
[1] Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf