Team:TU Delft/23 June 2010 content

From 2010.igem.org

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<h4>Characterization of Anderson RBS sequences</h4>  
<h4>Characterization of Anderson RBS sequences</h4>  
[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the overnight digested BioBricks. The following ligation reactions was performed:  
[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the overnight digested BioBricks. The following ligation reactions was performed:  
 +
1. I10341 + J1100 + pSB3C5
1. I10341 + J1100 + pSB3C5
 +
2. I10341 + J1101 + pSB3C5
2. I10341 + J1101 + pSB3C5
 +
3. I10341 + J1107 + pSB3C5
3. I10341 + J1107 + pSB3C5
 +
4. I10341 + J1117 + pSB3C5
4. I10341 + J1117 + pSB3C5
 +
5. I10341 + J1127 + pSB3C5
5. I10341 + J1127 + pSB3C5
 +
6. J1100 + pSB3C5 (negative control)
6. J1100 + pSB3C5 (negative control)

Revision as of 18:05, 17 July 2010

Lab Work

Characterization of Anderson RBS sequences

Ligations were performed using the overnight digested BioBricks. The following ligation reactions was performed:

1. I10341 + J1100 + pSB3C5

2. I10341 + J1101 + pSB3C5

3. I10341 + J1107 + pSB3C5

4. I10341 + J1117 + pSB3C5

5. I10341 + J1127 + pSB3C5

6. J1100 + pSB3C5 (negative control)

The general composition was:

Component Sample
I10341 3 μL
J611XX 4 μL
pSB3C5 1.5 μL
Ligation Buffer 3 μL
T4 ligase 1 μL
H20 15 μL

We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added. The mixtures were incubated overnight at 16 °C.