Team:KULeuven/30 August 2008

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  Input Output Filter InverTimer Reset Cell Death Memory

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  Introduction Three A's

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Lab Work

Wet Lab

A glycerol stock was made of the new parts and of our own parts ([http://partsregistry.org/Part:BBa_K145151 K145151], [http://partsregistry.org/Part:BBa_K145001 K145001], [http://partsregistry.org/Part:BBa_K145015 K145015]). We forgot one, so a liquid culture was made of part [http://partsregistry.org/Part:BBa_R0053 R0053].

We miniprepped the new parts and the two ligations that succeeded ([http://partsregistry.org/Part:BBa_C0040 C0040]+[http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_C0060 C0060]+[http://partsregistry.org/Part:BBa_B0015 B0015] that were plated on Ap instead of Km).

The digests with XbaI were started: [http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_B0033 B0033], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J23066 J23066], [http://partsregistry.org/Part:BBa_P0353 P0353], [http://partsregistry.org/Part:BBa_P0152 P0152], [http://partsregistry.org/Part:BBa_E0240 E0240].

The ligations we made yesterday were electroporated today (still in electrocompetent Top10 cells): [http://partsregistry.org/Part:BBa_R0011 R0011]+[http://partsregistry.org/Part:BBa_B0032 B0032], [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_K145001 K145001]+[http://partsregistry.org/Part:BBa_B0015 B0015], [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:BBa_B0015 B0015] and ([http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23022 J23022])+([http://partsregistry.org/Part:BBa_J23109 J23109]+[http://partsregistry.org/Part:BBa_J23032 J23032]).

Dry Lab

General

Created the oligos for the B1002 and B1006 terminators [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=terminator link]

Modeling

Final updated model uploaded to the wiki (only MatLab)

Remarks

I don't think the dam methylation is the problem with our ligations. This is the prefix: gaattcgcggccgcttctagag. The recognition site of XbaI is in bold. There is no overlap for dam methylation (gatc).

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