Team:Freiburg/Notebook/15 June

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==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
===PCR for Ccas and CcaR===
===PCR for Ccas and CcaR===
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==3A-assembly:Digestion==
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'''Investigators: Theo, Jakob'''
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Restriction digest of plasmid constructs (3A Assemblies PR1, PR2, PR3, PR4, PR5, PR7, PR8, Pr9)<br/>
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Protocol: Mix => for one reaction, total volume 10µl<br/>
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3µl DNA<br/>
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1µl NEBuffer4<br/>
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1µl BSA<br/>
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0,5µl Enzym<br/>
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<br/>
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==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
===Transformation===
===Transformation===
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==3A-assembly:Digestion==
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'''Investigators: Theo, Jakob'''
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+
-
Restriction digest of plasmid constructs (3A Assemblies PR1, PR2, PR3, PR4, PR5, PR7, PR8, Pr9)<br/>
+
-
Protocol: Mix => for one reaction, total volume 10µl<br/>
+
-
3µl DNA<br/>
+
-
1µl NEBuffer4<br/>
+
-
1µl BSA<br/>
+
-
0,5µl Enzym<br/>
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<br/>
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==<span style="color:green;">green light receptor</span==
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===Gel with PCR products: Ccas, Ccar===
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the first gel was stained with G-Stain, but adding G-Stain to the GeneRuler destroyed it.
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Second Gel was post-stained with ethidium-bromid, the bands for  CcaS &CcaR were at some place, nothing really clear.
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<span style= "color:blue;"> Comments: Has to be repeated. Either repeat the PCR or run a new gel with higher agarose concentration.</span>
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Latest revision as of 01:20, 22 September 2011


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

Contents

green light receptor

PCR for Ccas and CcaR

Investigators: Julia

Ccas: cyanobacteriochrome is a green light receptor that induces the expression of pycobillisome linker protein
Ccar: cognate response regulator

Primer diluted, PCR for CcaS with P(rimer) 5, P7 and CcaR with P4, P6

32,5 µl H2O
2µl Primer fw
2µl Primer rev
1µl DNA ( synechocystis from Hess-Lab)
1µl dNTPs
0,5 µl Phusion

Gel with PCR products: Ccas, Ccar

the first gel was stained with G-Stain, but adding G-Stain to the GeneRuler destroyed it. Second Gel was post-stained with ethidium-bromid, the bands for CcaS &CcaR were at some place, nothing really clear.

Comments: Has to be repeated. Either repeat the PCR or run a new gel with higher agarose concentration.

Miniprep

Investigators: Julia, Sandra

Comments: Miniprep from june,14 was showed bad results. Grew over night at 37°C.

Miniprep PR4a, PR4b, PR8a, PR8b, PR9a, PR9b
measurement of nanodrop was ok


3A-assembly:Digestion

Investigators: Theo, Jakob

Restriction digest of plasmid constructs (3A Assemblies PR1, PR2, PR3, PR4, PR5, PR7, PR8, Pr9)
Protocol: Mix => for one reaction, total volume 10µl
3µl DNA
1µl NEBuffer4
1µl BSA
0,5µl Enzym

Lysis cassette

Transformation

Investigators: Sandra

Transformation of BBa_K124014 bacteriophage Holin Gene pS105
plated out at 3:36pm in plates with ampicillin

Investigators: Julia

Transformation of BBa_E0040 green fluorescent protein. Ampicillin resistance.

                       

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