Team:TU Delft/23 June 2010 content

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[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the overnight [http://2010.igem.org/Team:TU_Delft#/blog?blog=22_June_2010 digested BioBricks]. The following ligation reactions was performed:  
[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the overnight [http://2010.igem.org/Team:TU_Delft#/blog?blog=22_June_2010 digested BioBricks]. The following ligation reactions was performed:  
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1. I10341 + J1100 + pSB3C5
 
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2. I10341 + J1101 + pSB3C5
 
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3. I10341 + J1107 + pSB3C5
 
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4. I10341 + J1117 + pSB3C5
 
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5. I10341 + J1127 + pSB3C5
 
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6. J1100 + pSB3C5 (negative control)
 
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The general composition was:
 
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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|'''Component'''
+
|'''#'''
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|'''Sample'''
+
|'''Ligation reaction'''
|-
|-
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|I10341
+
|1
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|3 μL
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|3 μL I10341 + 4 μL J1100 + 1.5 μL pSB3C5
|-
|-
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|J611XX
+
|2
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|4 μL
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|3 μL I10341 + 4 μL J1101 + 1.5 μL pSB3C5
|-
|-
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|pSB3C5
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|3
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|1.5 μL
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|3 μL I10341 + 4 μL J1107 + 1.5 μL pSB3C5
|-
|-
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|Ligation Buffer
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|4
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|3 μL
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|3 μL I10341 + 4 μL J1117 + 1.5 μL pSB3C5
|-
|-
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|T4 ligase
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|5
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|1 μL
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|3 μL I10341 + 4 μL J1127 + 1.5 μL pSB3C5
|-
|-
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|H20
+
|6
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|15 μL
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|4 μL J1100 + 1.5 μL pSB3C5 (negative control)
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|}
+
|}
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+
We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added.
We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added.
The mixtures were incubated overnight at 16 °C.
The mixtures were incubated overnight at 16 °C.

Revision as of 19:35, 17 July 2010

Lab work

Characterization of Anderson RBS sequences

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

# Ligation reaction
1 3 μL I10341 + 4 μL J1100 + 1.5 μL pSB3C5
2 3 μL I10341 + 4 μL J1101 + 1.5 μL pSB3C5
3 3 μL I10341 + 4 μL J1107 + 1.5 μL pSB3C5
4 3 μL I10341 + 4 μL J1117 + 1.5 μL pSB3C5
5 3 μL I10341 + 4 μL J1127 + 1.5 μL pSB3C5
6 4 μL J1100 + 1.5 μL pSB3C5 (negative control)

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