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From 2011.igem.org

Contents 1 green light receptor 1.1 PCR for Ccas and CcaR 1.2 Gel with PCR products: Ccas, Ccar 2 Miniprep 3 3A-assembly:Digestion 4 Lysis cassette 4.1 Transformation

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.