Protocols Ligation
From 2008.igem.org
Cocktail:
5.5ul d.s.H2O
1ul Backbone (keep on ice)
2ul Insert (keep on ice)
1ul Buffer (T4 DNA buffer)
0.5ul Ligase (T4 DNA ligase)
== Negative Control 1 ==
- Use 2mM Tris-Cl in place of insert
== Negative Control 2 ==
- Use 2mM Tris-Cl in place of backbone
== Control for PNK ==
- Use 2ul of d.s. H2O instead of insert
Let stand at room temperature for 20 minutes