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Team:SDU-Denmark/Protocols/Ligations
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Ligations
Protocol 1
Mix the following:
- 2uL 10x Ligase buffer
- 1uL T4 DNA ligase
- 2 or 4 uL cut backbone
- 5 or 10 uL cut PCR product
Leave at 17 degress C overnight.
Use for electroporation.
Protocol 2
Takes place in eppendorf tube.
- 2 ul 10x T4 ligase buffer
- 1 ul T4 ligase
- 5 ul PCR product (cut) of each brick which is to be ligated together - or 1 part plasmid and 5 part bricks
Leave at 17 degrees over-night.
Test ligation using PCR and run a test gel afterwards in order to check the PCR product has the right size.
