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.