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Team:KULeuven/20 August 2008
From 2008.igem.org
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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].
