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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=== | ||
Revision as of 00:45, 22 September 2011
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.
