Team:UCLA/Notebook/Spider Silk Genetics/27 April 2015
4/27/2015
1 2AB, 2BC, 2CA Sticky End Ligation Test
| AB+BC | BC+CA | CA+AB | AB+BC+CA | |
| 2x T7 Ligase Buffer | 5 uL | 5 uL | 5 uL | 5 uL |
| ddH2O | 1.5 uL | 1.5 uL | 0.5 uL | 0 uL |
| DNA 1 | 2 uL | 1 uL | 2 uL | 2 uL |
| DNA 2 | 1 uL | 2 uL | 2 uL | 1 uL |
| DNA 3 | n/a | n/a | n/a | 2 uL |
| T7 Ligase | 0.5 uL | 0.5 uL | 0.5 ul | 0.5 uL |
| Total | 10 uL | 10 uL | 10 uL | 10 uL |
| 25 C | 5 min |
| 65 C |
∙ The ligation test was run on 2% gel with 2 uL of 50 bp ladder. Results shown below.
Figure 1: T7 Ligation Test with MaSp2 fragments. BC was used as a
negative ligation control. The presence of ladders may indicate non-specific
binding.