From 2007.igem.org

Information obtained from:

Promega

We used one ligase for the ligations:

• T4 DNA Ligase

Protocol obtained from: Promega

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Ligation

Usually we ligate following the recommendations from the commercial information:

we have had good results with a molar ratio of 3:1

• Ligase

total DNA	20-200 ng
enzyme	1 uL
2X Rapid Ligation buffer	10 uL
ddwater	up to 20 uL (sometimes, if the samples are in low concentrations, we go up to 40uL)