Team:Freiburg/Notebook/12 September

From 2011.igem.org

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<div id="notebook-page-header">
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<div id="notebook-back" width="100px" >
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<a href="https://2011.igem.org/Team:Freiburg/Notebook/11_September">Previous entry</a>
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</div>
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<div id="notebook-title">
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<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 12 September </a>
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</div>
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<div id="notebook-next">
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<a href="https://2011.igem.org/Team:Freiburg/Notebook/13_September">Next entry</a>
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</html>
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==Commons==
 +
 +
===Minipreps===
 +
 +
'''Investigators: Sandra'''
 +
 +
Because of the positive testdigest, a miniprep of the PR+GFP was performed. Samples were send for sequencing.
 +
==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
Line 7: Line 32:
'''Investigators:NAME'''
'''Investigators:NAME'''
 +
==<span style="color:blue;">blue light receptor</span>==
 +
===Miniprep of PR+lovtap-notgate domain===
-
==<span style="color:blue;">blue light receptor</span>==
+
'''Investigators: Sandra'''
-
===NAME OF YOUR EXPERIMENT===
+
Miniprep of:
 +
*39A
 +
*39B
 +
*40A
 +
*40B
 +
*41A
 +
*41B
 +
*42A
 +
*42B
 +
*44A
 +
*44B
-
'''Investigators:NAME'''
+
39=PR1,
 +
40=PR2,
 +
41=PR3,
 +
42=PR4,
 +
44=PR6.
 +
===2A-assembly: Digestion===
 +
'''Investigators: Sandra'''
 +
 +
Digestion of the samples from miniprep (10 samples) and digestion of RFP (S1b, S2b).
 +
 +
'''PR with Lov-Notgate domain:'''
 +
 +
*39A
 +
*39B
 +
*40A
 +
*40B
 +
*41A
 +
*41B
 +
*42A
 +
*42B
 +
*44A
 +
*44B
 +
 +
digested with NehI and PstI.
 +
 +
'''RFP:'''
 +
 +
*S1b
 +
*S2b
 +
 +
digested with SpeI and PstI.
 +
 +
 +
===2A-assembly: Ligation===
 +
 +
Ligation of RFP with 39-44.
 +
 +
*39A-S1b
 +
*39A-S2b
 +
 +
*39B-S1b
 +
*39B-S2b
 +
 +
 +
*40A-S1b
 +
*40A-S2b
 +
 +
*40B-S1b
 +
*40B-S2b
 +
 +
 +
*41A-S1b
 +
*41A-S2b
 +
 +
*41B-S1b
 +
*41B-S2b
 +
 +
 +
*42A-S1b
 +
*42A-S2b
 +
 +
*42B-S1b
 +
*42B-S2b
 +
 +
 +
*44A-S1b
 +
*44A-S2b
 +
 +
*44B-S1b
 +
*44B-S2b
 +
<br/>
 +
 +
===Trafo===
 +
 +
 +
Trafo of the ligated samples on plates with Cm.
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==
Line 29: Line 141:
==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
-
===NAME OF YOUR EXPERIMENT===
+
===High throughput sequencing===
-
'''Investigators:NAME'''
+
'''Investigators:Theo'''
 +
All PR parts in C3 vectors, M48 in C3 and also the M48+1 and M48+7 in Amp vectors were sent for sequencing
==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==
 +
 +
Testdigest
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name:Rüdiger
 +
 +
 +
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 12.09.
 +
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Experiment Miniprep (Date) 10.&11.09.
 +
 +
(Name) Rüdiger
 +
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name: Precipitator
 +
 +
|}
 +
For one reaction you need: For Mastermix: Number of samples+2extra
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 4μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>O
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 280
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Buffer, NEB4
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 70
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| BSA (10x)
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 70
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 0,5 μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Enzym 1
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 35
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Eco R1
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 0,5 μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Enzym 2
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 3 μl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|}
 +
10 μl total volume
 +
 +
 +
Give 3 μl of DNA in an eppi and add 7μl of the mastermix.
 +
 +
Incubate for about 1h at 37°C.
 +
 +
 +
Add 1 μl Loading dye buffer and load the gel.
 +
 +
Take a picture of the gel, print picture and label the lanes!
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1aB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1aC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1bB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 1bC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 2aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 2aB
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 2aC
 +
 +
|-
 +
| colspan="9"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Bam H1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| X
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 2bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 2bB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 2bC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 3bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 3bB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 3bC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 3aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 3aB
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 3aC
 +
 +
|-
 +
| colspan="9"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Bam H1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| X
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 6aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 6aB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 6aC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 74aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 7aB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 7aC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 7bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 7bB
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 7bC
 +
 +
|-
 +
| colspan="9"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Pst 1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| X
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5bB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5bC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5aA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5aB
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 5aC
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 6bA
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 6bB
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 6bC
 +
 +
|-
 +
| colspan="9"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Pst 1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| X
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8a1
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8a3
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8a4
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8b1
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8b2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 8b4
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10a2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10a3
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 10a4
 +
 +
|-
 +
| colspan="9"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Spc 1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| X
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9a1
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9a2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9a3
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9a4
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9b1
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9b2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 9b3
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 9b4
 +
 +
|-
 +
| colspan="8"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Spc 1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| V
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| V
 +
 +
|}
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10b3
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10b4
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10b01
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 10b2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 4a1
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 4a2
 +
| style="border-top:0.0007in solid #000000;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| 4a3
 +
| style="border:0.0007in solid #000000;padding:0.0382in;"| 4b4
 +
 +
|-
 +
| colspan="8"  style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| Eco R1 + Spc 1
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| V
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| V
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| V
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| X
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:none;padding:0.0382in;"| V
 +
| style="border-top:none;border-bottom:0.0007in solid #000000;border-left:0.0007in solid #000000;border-right:0.0007in solid #000000;padding:0.0382in;"| V
 +
 +
|}
 +
X=Failure, V=Success
 +
===NAME OF YOUR EXPERIMENT===
===NAME OF YOUR EXPERIMENT===

Latest revision as of 00:15, 22 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

Commons

Minipreps

Investigators: Sandra

Because of the positive testdigest, a miniprep of the PR+GFP was performed. Samples were send for sequencing.


green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

blue light receptor

Miniprep of PR+lovtap-notgate domain

Investigators: Sandra

Miniprep of:

  • 39A
  • 39B
  • 40A
  • 40B
  • 41A
  • 41B
  • 42A
  • 42B
  • 44A
  • 44B

39=PR1, 40=PR2, 41=PR3, 42=PR4, 44=PR6.

2A-assembly: Digestion

Investigators: Sandra


Digestion of the samples from miniprep (10 samples) and digestion of RFP (S1b, S2b).

PR with Lov-Notgate domain:

  • 39A
  • 39B
  • 40A
  • 40B
  • 41A
  • 41B
  • 42A
  • 42B
  • 44A
  • 44B

digested with NehI and PstI.

RFP:

  • S1b
  • S2b

digested with SpeI and PstI.


2A-assembly: Ligation

Ligation of RFP with 39-44.

  • 39A-S1b
  • 39A-S2b
  • 39B-S1b
  • 39B-S2b


  • 40A-S1b
  • 40A-S2b
  • 40B-S1b
  • 40B-S2b


  • 41A-S1b
  • 41A-S2b
  • 41B-S1b
  • 41B-S2b


  • 42A-S1b
  • 42A-S2b
  • 42B-S1b
  • 42B-S2b


  • 44A-S1b
  • 44A-S2b
  • 44B-S1b
  • 44B-S2b


Trafo

Trafo of the ligated samples on plates with Cm.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

High throughput sequencing

Investigators:Theo


All PR parts in C3 vectors, M48 in C3 and also the M48+1 and M48+7 in Amp vectors were sent for sequencing

Precipitator

Testdigest


Name:Rüdiger


Date: 12.09.
Continue from Experiment Miniprep (Date) 10.&11.09.

(Name) Rüdiger

Project Name: Precipitator

For one reaction you need: For Mastermix: Number of samples+2extra


4μl H2O 280
1μl Buffer, NEB4 70
1μl BSA (10x) 70
0,5 μl Enzym 1 35 Eco R1
0,5 μl Enzym 2
3 μl DNA

10 μl total volume


Give 3 μl of DNA in an eppi and add 7μl of the mastermix.

Incubate for about 1h at 37°C.


Add 1 μl Loading dye buffer and load the gel.

Take a picture of the gel, print picture and label the lanes!


1aA 1aB 1aC 1bA 1bB 1bC 2aA 2aB 2aC
Eco R1 + Bam H1
X X X X X X X X X
2bA 2bB 2bC 3bA 3bB 3bC 3aA 3aB 3aC
Eco R1 + Bam H1
X X X X X X X X X
6aA 6aB 6aC 74aA 7aB 7aC 7bA 7bB 7bC
Eco R1 + Pst 1
X X X X X X X X X
5bA 5bB 5bC 5aA 5aB 5aC 6bA 6bB 6bC
Eco R1 + Pst 1
X X X X X X X X X
8a1 8a3 8a4 8b1 8b2 8b4 10a2 10a3 10a4
Eco R1 + Spc 1
X X X X X X X X X
9a1 9a2 9a3 9a4 9b1 9b2 9b3 9b4
Eco R1 + Spc 1
X X X X V X X V
10b3 10b4 10b01 10b2 4a1 4a2 4a3 4b4
Eco R1 + Spc 1
V V X V X X V V

X=Failure, V=Success


NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/12_September"