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Team:KULeuven/27 August 2008
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== Lab Work == | == Lab Work == | ||
=== Wet Lab === | === Wet Lab === | ||
| - | + | We continued our ligations. Therefore we miniprepped and digested the following parts: [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], [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] (with ''Xba''I) and [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23022 J23022] (with ''Eco''RI and ''Spe''I). However, when we put the digests on an agarose gel, we didn't see anything. But, according to the nanodrop, there should be DNA in our samples. So we put the miniprepped plasmids on gel (not digested) and we couldn't see anything - again. This means that either the miniprep failed or the colonies were crap. Tomorrow we will electroporate the ligated parts again (but we will plate them on Ap instead of Km). We will also redo the ligations, but this time we will use the protein coding regions as vector and B0015 as insert. This means cutting the vectors with ''Spe''I and ''Pst''I and B0015 with ''Xba''I and ''Pst''I. | |
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| - | + | Modelling has shown that our original setup requires some (major) adaptations. We miniprepped and digested some of the parts needed for these changes: [http://partsregistry.org/Part:BBa_B0033 B0033], [http://partsregistry.org/Part:BBa_B0034 B0034] and [http://partsregistry.org/Part:BBa_C0012 C0012]. We also did this for [http://partsregistry.org/Part:BBa_R0011 R0011] and [http://partsregistry.org/Part:BBa_F1610 F1610], but F1610 seemed to be degraded - again. | |
| - | + | There were colonies on the plates with [http://partsregistry.org/Part:BBa_K145001 K145001]+[http://partsregistry.org/Part:pSB1A2 pSB1A2] and [http://partsregistry.org/Part:BBa_K145151 K145151]+[http://partsregistry.org/Part:pSB1A2 pSB1A2]. Liquid cultures were made. | |
| - | + | We will make a glycerol stock of the ligations that succeeded. Therefore, we made a liquid culture of [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_I712074 I712074]+[http://partsregistry.org/Part:BBa_J23032 J23032], [http://partsregistry.org/Part:BBa_J23109 J23109]+[http://partsregistry.org/Part:BBa_J23032 J23032]and [http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_J23022 J23022]. | |
| - | ==== | + | === Dry Lab === |
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| - | + | ==== Ethics ==== | |
| + | Worked some more on the ethics-part. Did some more research. | ||
Latest revision as of 11:54, 12 October 2008
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Contents |
Lab Work
Wet Lab
We continued our ligations. Therefore we miniprepped and digested the following parts: C0040+B0015, C0012+B0015, K145015+B0015, K145001+B0015, K145151+B0015 (with XbaI) and R0040+J23022 (with EcoRI and SpeI). However, when we put the digests on an agarose gel, we didn't see anything. But, according to the nanodrop, there should be DNA in our samples. So we put the miniprepped plasmids on gel (not digested) and we couldn't see anything - again. This means that either the miniprep failed or the colonies were crap. Tomorrow we will electroporate the ligated parts again (but we will plate them on Ap instead of Km). We will also redo the ligations, but this time we will use the protein coding regions as vector and B0015 as insert. This means cutting the vectors with SpeI and PstI and B0015 with XbaI and PstI.
Modelling has shown that our original setup requires some (major) adaptations. We miniprepped and digested some of the parts needed for these changes: B0033, B0034 and C0012. We also did this for R0011 and F1610, but F1610 seemed to be degraded - again.
There were colonies on the plates with K145001+pSB1A2 and K145151+pSB1A2. Liquid cultures were made.
We will make a glycerol stock of the ligations that succeeded. Therefore, we made a liquid culture of J23116+B0032, R0040+B0032, I712074+J23032, J23109+J23032and R0040+J23022.
Dry Lab
Ethics
Worked some more on the ethics-part. Did some more research.
