Team:SDU-Denmark/Protocols/Ligations

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Ligations

Protocol 1

Mix the following:

  1. 2uL 10x Ligase buffer
  2. 1uL T4 DNA ligase
  3. 2 or 4 uL cut backbone
  4. 5 or 10 uL cut PCR product

Leave at 17 degress C overnight.

Use for electroporation.


Protocol 2

Takes place in eppendorf tube.

  1. 2 ul 10x T4 ligase buffer
  2. 1 ul T4 ligase
  3. 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.