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Team:SDU-Denmark/Protocols/Ligations
From 2009.igem.org
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=Ligations= | =Ligations= | ||
| - | + | =Protocol 1= | |
Mix the following: | Mix the following: | ||
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Use for electroporation. | Use for electroporation. | ||
| + | |||
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| + | =Protocol 2= | ||
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| + | 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. | ||
Latest revision as of 10:11, 17 August 2009
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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.
