Team:Freiburg/Notebook/5 August
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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/4_August">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 5 August </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/6_August">Next entry</a> | ||
+ | </div> | ||
+ | </div> | ||
+ | </html> | ||
+ | |||
==Meeting== | ==Meeting== | ||
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===other stuff=== | ===other stuff=== | ||
*start creating nice pictures to explain your subproject and write text to explain the pictures | *start creating nice pictures to explain your subproject and write text to explain the pictures | ||
- | *Sandra organized Freehand from the MPI, it is installed | + | *Sandra organized Freehand from the MPI, it is installed on both mac books |
*give-aways: digital postcard, real postcard, fluorescent protein | *give-aways: digital postcard, real postcard, fluorescent protein | ||
*Tobi will ask at the financial department about the croud funding money | *Tobi will ask at the financial department about the croud funding money | ||
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==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
- | ===Quickchange=== | + | ===1.Quickchange=== |
'''Investigators: Jakob''' | '''Investigators: Jakob''' | ||
<br/> | <br/> | ||
- | + | ||
+ | ====PCR==== | ||
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|} | |} | ||
- | + | Program I used | |
{| style="border-spacing:0;" | {| style="border-spacing:0;" | ||
- | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 95 °C – 5’ | + | | rowspan="3" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 95 °C – 5’ |
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 95°C – 15 ‘’ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 95°C – 15 ‘’ | ||
- | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | + | | rowspan="3" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| x 20 |
- | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 72°C – 5’ | + | | rowspan="3" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 72°C – 5’ |
- | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 4°C – ∞ | + | | rowspan="3" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 4°C – ∞ |
|- | |- | ||
+ | |||
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 55°C – 15 ‘’ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 55°C – 15 ‘’ | ||
|- | |- | ||
+ | |||
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 72°C – 1,15’ + 2’’/Cycle | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 72°C – 1,15’ + 2’’/Cycle | ||
|} | |} | ||
- | |||
- | + | * Labeled as Jakob1 and Jakob2 in the fridge | |
+ | |||
+ | ==='''2'''.Digest quickchange PCR with Dpn1<br/>=== | ||
+ | |||
+ | |||
+ | 5µl Buffer NEB4 <br/> | ||
+ | 5µl BSA (10x) <br/> | ||
+ | 2µl Dpn1 <br/> | ||
+ | 38µl DNA from quickchange (CcaS) <br/> | ||
+ | incubate at 37°C for 1,5 h <br/> | ||
+ | inactivate at 80°C for 20 min. <br/> | ||
+ | <br/> | ||
+ | * To-do: Transformation | ||
+ | |||
+ | ===<span style="color:#0000EE"> '''3''' </span><span style="color:#00EEEE"> . </span><span style="color:#4EEE94"> T </span><span style="color:#00EE00"> r </span><span style="color:#BCEE68"> a </span><span style="color:#FFFF00"> n </span><span style="color:#FFB90F"> s </span><span style="color:#FF3030"> f </span><span style="color:#FF4500"> o </span><span style="color:#CD0000"> r </span><span style="color:#FF1493"> m </span><span style="color:#FF00FF"> a </span><span style="color:#E066FF"> t </span><span style="color:#9B30FF"> i </span><span style="color:#AB82FF"> o </span><span style="color:#FFE1FF"> n </span><br/>=== | ||
+ | |||
+ | ==='''4'''.Picked some colonies<br/>=== | ||
+ | from transformation yesterday | ||
- | + | *To-do: pick colonies, do Miniprep | |
==<span style="color:blue;">blue light receptor</span>== | ==<span style="color:blue;">blue light receptor</span>== | ||
- | === | + | ===Gradient PCR of Lovtap with new primer=== |
- | '''Investigators: | + | '''Investigators: Sandra''' |
+ | Primer:<br/> | ||
+ | * P38 (former name:P36) LOVdw<br/> | ||
+ | * P42 LOVtap up<br/> | ||
+ | DNA template:<br/> | ||
+ | * M35 (Lovtap)<br/> | ||
+ | |||
+ | |||
+ | PCR program: <br/> | ||
+ | * "Lovtap ohne Überhänge" with temperature gradient during the annealing step from 50°C-60°C. The elongation step and the final elongation step were extended. | ||
+ | |||
+ | "cold":50°C<br/> | ||
+ | "warm":60°C<br/> | ||
+ | [[File:Freiburg_2011_LovTAP.jpg|caption|border|300px]] | ||
+ | |||
+ | Comments: PCR did not work again. We will try to do 3A-assembly. | ||
+ | <br/> | ||
+ | <br/> | ||
==<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>== | ||
- | === | + | ===Lysis Cassette +RBS=== |
- | + | ||
- | + | ||
- | + | ||
+ | '''Investigators:Theo''' | ||
+ | No colonies, repeated transformation as August 4th | ||
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== |
Latest revision as of 01:05, 22 September 2011