Team:TU Delft/23 June 2010 content

From 2010.igem.org

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===Lab Work===
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==Lab Work==
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[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the digested BioBricks obtained in the early morning. We following ligation was performed:
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<h4>Characterization of Anderson RBS sequences</h4>
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[[Team:TU_Delft/protocols/ligation|Ligations]] were performed using the overnight 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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|Componet
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|'''Component'''
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|Sample
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|'''Sample'''
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|I10341
|I10341
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|15 μL
|15 μL
|}
|}
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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.
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The negative control for the ligation consisted of J61100 + pSB3C5. The mixtures were incubated overnight at 16 degrees.
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The mixtures were incubated overnight at 16 °C.

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.