Team:Freiburg/Notebook/2 September
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+ | <div id="notebook-page-header"> | ||
+ | <div id="notebook-back" width="100px" > | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/1_September">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 2 September </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/3_September">Next entry</a> | ||
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==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
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Miniprep of new ligated parts in pSB1C3 vector. | Miniprep of new ligated parts in pSB1C3 vector. | ||
- | After miniprep samples were digested and loaded onto a gel. | + | After miniprep samples were digested and loaded onto a gel. Not c has inserts:-) |
[[File:Freiburg2011_Lovtap.jpg|400px]] | [[File:Freiburg2011_Lovtap.jpg|400px]] | ||
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+ | [[File:Freiburg2011Lovtap_and_greenlight.jpg|400px]] | ||
+ | |||
+ | 1:2 not a has inserts too! | ||
+ | |||
+ | ===Digestion: 2A-assembly=== | ||
+ | |||
+ | '''Investigators: Sophie''' | ||
+ | |||
+ | Digestion of the PCR products Notgate, Lovtap for the 2A-assembly. | ||
+ | |||
+ | ===Ligation:2A-assembly=== | ||
+ | |||
+ | '''Investigators: Sandra''' | ||
+ | |||
+ | Ligation of the digested parts in the already digested vector pSB1C3. | ||
+ | Ligation at room temperature over night. | ||
==<span style="color:red;">red light receptor</span>== | ==<span style="color:red;">red light receptor</span>== | ||
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==<span style="color:orange;">Lysis cassette</span>== | ==<span style="color:orange;">Lysis cassette</span>== | ||
- | === | + | ===3A Assembly of Quickchange-modified K098995 + modified Lysis genes K124017=== |
- | '''Investigators: | + | '''Investigators:Theo''' |
+ | <br> | ||
+ | Sequencing of Part S66 (modified Lysis genes K124017) showed that it is a DNA fragment from our other projects, so another miniprep was done and sent for sequencing. | ||
+ | *Miniprep data were good: | ||
+ | **Concentration = 220ng/mikrol | ||
+ | **260/280 = 2.04 | ||
+ | **260/230 = 2.19 | ||
+ | <br> | ||
+ | No gel was run to see if there is a band! | ||
+ | <br> | ||
+ | <br/> | ||
+ | ==<span style="color:grey;">Precipitator</span>== | ||
+ | '''Rüdiger''' 02.-05.09. | ||
+ | |||
+ | |||
+ | Trafo, colony picking & miniprep of ligations from 01.09. | ||
+ | |||
+ | Nothing worked | ||
+ | |||
+ | Another trafo done with same ligation. Found out that linearized C3 vector has no SpC1 restriction site. | ||
- | |||
===Trafo=== | ===Trafo=== | ||
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#pSB1C3 | #pSB1C3 | ||
#pSB1C3 | #pSB1C3 | ||
+ | |||
+ | ===Miniprep=== | ||
+ | |||
+ | '''Investigators: Sophie''' | ||
+ | |||
+ | miniprep of GFP-pbd with IPTG-inducible promoter. The pellets were green!!! So we finally got both promoter and plastic binding domain into a vector! We can make experiments with the pbd now. | ||
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Latest revision as of 00:27, 22 September 2011