Team:KULeuven/20 August 2008

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=== Wet Lab ===
=== Wet Lab ===
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* Ligations C0060+B0015, C0040+B0015, C0012+B0015 and K145151+B0015 were miniprepped, but were mixed up, so have to be redone. They are cut with ''Xba''I anyway. Colonies were re-inoculated in liquid medium to grow overnight and to be miniprepped again tomorrow.
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Ligations [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015] were miniprepped, but were mixed up, so have to be redone. They were cut with ''Xba''I anyway, but we couldn't see anything on the gel (??). Colonies were re-inoculated in liquid medium to grow overnight and to be miniprepped again tomorrow.
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* We tried to put K145001 (T7 polymerase without tag) into a vector using TOPO TA cloning.
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* We performed a PCR on the Klenow mix of the end-filling (part K145150). There were no bands, so it failed. We also tried to digest this mixture and ligate it into a pSB1A2 vector (without purifying the mixtures first). Hopefully we'll have colonies tomorrow.
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* We set up some ligations: J23116+B0032, R0040+B0032, R0011+F1610 and K145151+pSB1A2.
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=== Dry Lab ===
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We tried to put [http://partsregistry.org/Part:BBa_K145001 K145001] (T7 polymerase without tag) into a vector using TOPO TA cloning. We did this because the previous attempts to put this part into a pSB1A2 vector failed.
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==== Modeling ====
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We performed a PCR on the Klenow mix of the end-filling (part [http://partsregistry.org/Part:BBa_K145150 K145150]) with primers 1129 and 1130. There were no bands, so it failed. We also tried to digest this mixture and ligate it into a pSB1A2 vector (without purifying the mixtures first). Hopefully we'll have colonies tomorrow.
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==== Wiki ====
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We set up some ligations: [http://partsregistry.org/Part:BBa_J23116 J23116]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0011 R0011]+[http://partsregistry.org/Part:BBa_F1610 F1610] and [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:pSB1A2 pSB1A2].
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== Remarks ==
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Latest revision as of 16:47, 11 October 2008

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Lab Work

Wet Lab

Ligations [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_C0012 C0012]+[http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_K145015 K145015]+[http://partsregistry.org/Part:BBa_B0015 B0015] were miniprepped, but were mixed up, so have to be redone. They were cut with XbaI anyway, but we couldn't see anything on the gel (??). Colonies were re-inoculated in liquid medium to grow overnight and to be miniprepped again tomorrow.

We tried to put [http://partsregistry.org/Part:BBa_K145001 K145001] (T7 polymerase without tag) into a vector using TOPO TA cloning. We did this because the previous attempts to put this part into a pSB1A2 vector failed.

We performed a PCR on the Klenow mix of the end-filling (part [http://partsregistry.org/Part:BBa_K145150 K145150]) with primers 1129 and 1130. There were no bands, so it failed. We also tried to digest this mixture and ligate it into a pSB1A2 vector (without purifying the mixtures first). Hopefully we'll have colonies tomorrow.

We set up some ligations: [http://partsregistry.org/Part:BBa_J23116 J23116]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_R0011 R0011]+[http://partsregistry.org/Part:BBa_F1610 F1610] and [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:pSB1A2 pSB1A2].