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Team:KULeuven/31 July 2008
From 2008.igem.org
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== Lab Work == | == Lab Work == | ||
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=== Wet Lab === | === Wet Lab === | ||
| - | + | A gel was run with the parts that were cut with ''Xba''I during the night. [http://partsregistry.org/Part:BBa_J23022 J23022] was not cut, but fortunately [http://partsregistry.org/Part:BBa_B0032 B0032], [http://partsregistry.org/Part:BBa_B0015 B0015] and [http://partsregistry.org/Part:BBa_J23032 J23032] were cut. These positive parts were then cut with ''Eco''RI and were again separated on gel, followed by gel extraction. After that, we could perform the following ligations: [http://partsregistry.org/Part:BBa_J23100 J23100]+[http://partsregistry.org/Part:BBa_B0032 B0032] and [http://partsregistry.org/Part:BBa_R0084 R0084]+[http://partsregistry.org/Part:BBa_B0032 B0032]. | |
| - | + | We did a miniprep of the following parts: [http://partsregistry.org/Part:BBa_R0011 R0011], [http://partsregistry.org/Part:BBa_P1010 P1010], [http://partsregistry.org/Part:BBa_C0062 C0062], [http://partsregistry.org/Part:BBa_I712022 I712022], [http://partsregistry.org/Part:BBa_C0060 C0060] and [http://partsregistry.org/Part:BBa_R0062 R0062]. We also nanodropped them and digested them with ''Spe''I. The ''Spe''I of La Roche is empty, so we used the enzyme of NEB. Because ''Eco''RI doesn't cut very well in the buffer of ''Spe''I (NEB buffer 2), we began to purify it by ethanol-precipitation. We put it overnight in -20°C. | |
| - | + | Competent TOP10 cells were transformed by heat shock with the overnight ligation products. | |
=== Dry Lab === | === Dry Lab === | ||
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| - | == | + | ==== General ==== |
| - | + | Primers for EnvZ::KmR mutation were made, will be ordered tomorrow. | |
| - | + | ==== Wiki ==== | |
| - | + | Worked on count down, keeping track on the time distance till the jamboree. Timer will be done tomorrow, after that the homepage will be taken care of. | |
| - | + | "The Project" tab has been taken care of, adding the details/mechanisms of the different parts. Tried to add figures and schemes for easier reading. Will be worked on whenever I further feel like it. | |
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Revision as of 16:08, 18 September 2008
| << return to notebook | return to homepage >> | ||
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Contents |
Lab Work
Wet Lab
A gel was run with the parts that were cut with XbaI during the night. J23022 was not cut, but fortunately B0032, B0015 and J23032 were cut. These positive parts were then cut with EcoRI and were again separated on gel, followed by gel extraction. After that, we could perform the following ligations: J23100+B0032 and R0084+B0032.
We did a miniprep of the following parts: R0011, P1010, C0062, I712022, C0060 and R0062. We also nanodropped them and digested them with SpeI. The SpeI of La Roche is empty, so we used the enzyme of NEB. Because EcoRI doesn't cut very well in the buffer of SpeI (NEB buffer 2), we began to purify it by ethanol-precipitation. We put it overnight in -20°C.
Competent TOP10 cells were transformed by heat shock with the overnight ligation products.
Dry Lab
General
Primers for EnvZ::KmR mutation were made, will be ordered tomorrow.
Wiki
Worked on count down, keeping track on the time distance till the jamboree. Timer will be done tomorrow, after that the homepage will be taken care of.
"The Project" tab has been taken care of, adding the details/mechanisms of the different parts. Tried to add figures and schemes for easier reading. Will be worked on whenever I further feel like it.
