Team:KULeuven/2 September 2008

From 2008.igem.org

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=== Wet Lab ===
=== Wet Lab ===
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* We tried to construct our parts K145150 (hybrid promoter) and K145014 (T7 polymerase with UmuD tag) using PCR. Both of them failed, but K145150 gave two bands. This is probably because we didn't purify it at first. We will do that tomorrow.
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We tried to construct our parts [http://partsregistry.org/Part:BBa_K145150 K145150] (hybrid promoter) and [http://partsregistry.org/Part:BBa_K145014 K145014] (T7 polymerase with UmuD tag) using PCR. Both of them failed, but K145150 gave two bands. This is probably because we didn't purify it at first. We will do that tomorrow.
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* We made electrocompetent cells of the JM109 stem.
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* We did some more digests: <div style="margin-left:10px;"> cut with ''Spe''I and ''Eco''RI -> R0040, B0014, R0053 and R1052 (R1052 failed). </div> <div style="margin-left:10px;"> cut with ''Xba''I -> K145151+B0015, C0040+B0015, C0060+B0015, C0012+B0015 and K145015+B0015. </div>
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* We started a few ligations: R0040+B0033, R0053+P0152, R0040+J23066, B0014+B0033 and R0040+E0240.
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* We also tested the ligations of 31 August using PCR. Most of them were OK. The ligations that succeeded so far can be found on the schemes below.
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We made electrocompetent cells of the JM109 stem.
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<div style="margin-left:100px;"> Current Status (Final Scheme): </div>
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We did some more digests:  
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[[Image:2sept final.PNG|center]]
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* cut with ''Spe''I and ''Eco''RI -> [http://partsregistry.org/Part:BBa_R0040 R0040], [http://partsregistry.org/Part:BBa_B0014 B0014], [http://partsregistry.org/Part:BBa_R0053 R0053] and [http://partsregistry.org/Part:BBa_R1052 R1052](R1052 failed).
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<div style="margin-left:100px;"> Current Status (Parallel Scheme): </div>
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* cut with ''Xba''I -> [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:BBa_B0015 B0015].
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[[Image:2sept parallel.PNG|center]]
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We started a few ligations: [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0033 B0033], [http://partsregistry.org/Part:BBa_R0053 R0053]+[http://partsregistry.org/Part:BBa_P1052 P1052], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23066 J23066], [http://partsregistry.org/Part:BBa_B0014 B0014]+[http://partsregistry.org/Part:BBa_B0033 B0033] and [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_E0240 E0240].
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We also tested the ligations of 31 August using PCR. Most of them were OK. The ligations that succeeded so far can be found on the schemes below.
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[[Image:2sept final.PNG|left|thumb|370px|Current Status (Final Scheme)]]
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[[Image:2sept parallel.PNG|center|thumb|387px|Current Status (Parallel Scheme)]]
=== Dry Lab ===
=== Dry Lab ===

Latest revision as of 01:04, 30 October 2008

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Contents

Lab Work

Wet Lab

We tried to construct our parts [http://partsregistry.org/Part:BBa_K145150 K145150] (hybrid promoter) and [http://partsregistry.org/Part:BBa_K145014 K145014] (T7 polymerase with UmuD tag) using PCR. Both of them failed, but K145150 gave two bands. This is probably because we didn't purify it at first. We will do that tomorrow.

We made electrocompetent cells of the JM109 stem.

We did some more digests:

  • cut with SpeI and EcoRI -> [http://partsregistry.org/Part:BBa_R0040 R0040], [http://partsregistry.org/Part:BBa_B0014 B0014], [http://partsregistry.org/Part:BBa_R0053 R0053] and [http://partsregistry.org/Part:BBa_R1052 R1052](R1052 failed).
  • cut with XbaI -> [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:BBa_B0015 B0015].

We started a few ligations: [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0033 B0033], [http://partsregistry.org/Part:BBa_R0053 R0053]+[http://partsregistry.org/Part:BBa_P1052 P1052], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23066 J23066], [http://partsregistry.org/Part:BBa_B0014 B0014]+[http://partsregistry.org/Part:BBa_B0033 B0033] and [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_E0240 E0240].

We also tested the ligations of 31 August using PCR. Most of them were OK. The ligations that succeeded so far can be found on the schemes below.

Current Status (Final Scheme)
Current Status (Parallel Scheme)

Dry Lab

Ethics...

Modeling

Latex-tool... Multi-cell... Diffusion... Updating wiki...

Wiki

Remarks