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Team:Freiburg/Notebook/2 September
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{{:Team:Freiburg/Templates/header}} | {{:Team:Freiburg/Templates/header}} | ||
| - | + | <html> | |
| + | <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> | ||
| + | </div> | ||
| + | </div> | ||
| + | </html> | ||
==<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]] | + | [[File:Freiburg2011_Lovtap.jpg|400px]] |
| + | |||
| + | [[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>== | ||
| Line 32: | Line 61: | ||
==<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=== | ||
| Line 56: | Line 103: | ||
#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. | ||
{{:Team:Freiburg/Templates/footer}} | {{:Team:Freiburg/Templates/footer}} | ||
Latest revision as of 00:27, 22 September 2011
Contact 
Contents |
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
MIniprep
Investigators: Sophie, Sandra
Miniprep of new ligated parts in pSB1C3 vector.
After miniprep samples were digested and loaded onto a gel. Not c has inserts:-)
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.
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
3A Assembly of Quickchange-modified K098995 + modified Lysis genes K124017
Investigators:Theo
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
No gel was run to see if there is a band!
Precipitator
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
Investigators: Julia
Trafo of ligated parts:
- pGEX vector (Amp. resistance)
- pGEX vector (Amp. resistance)
- pGEX vector (Amp. resistance)
- pSB1C3
- pET Duet vector (Amp. resistance)
- pET Duet vector (Amp. resistance)
- pET Duet vector (Amp. resistance)
- 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.
