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From 2010.igem.org

Contents 1 Nina 2 Glycerol stock on CPP and IgG protease 3 Streaking out tyrosinase 4 PCR on tyrosinase 5 Agarose gel on tyrosinase and IgG protease

Nina

Glycerol stock on CPP and IgG protease

I made glycerol stocks on the CPP transportan 10 C version from dish +8ng/ul. Colony number 9 and 40 were added each to a falcon tube containing 12 ml LB and 24 ul ampicillin resistance (50 mg/ml). The tubes were incubated in 37 °C until OD reaches around 0.6. After around 4 hours the OD was about 0.6.

I also made a glycerol stock on IgG protease inserted in the linjerized shipping vector (with chloramphenicol resistance). Colony number 2 was added to a falcon tube containing 12 ml LB and 24 ul chloramphenicol resistance (50 mg/ml). The tubes were incubated in 37 °C until OD reaches around 0.6, which was obtained after around 5 hours.

Glycerol stock:

• 400 ul Glycerol

• 800 ul Bacterial sample

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Streaking out tyrosinase

I stroke out tyrosinase which we obatined from iGEM hq in a glycerol stock on a dish containing ampicillin resistance. It was incubated in 37 °C ON. This will be followed tomorrow with a colony-PCR.

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PCR on tyrosinase

I ran a PCR on tyrosinase (1ul) from a glycerol stock with the gene's forward and reverse primers. The annealing time was 2 min.

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Agarose gel on tyrosinase and IgG protease

Both the PCR product of tyrosinase from todays work and the mini prep of IgG protease in the shipping vector from yesterday's work were run on an agarose gel (2%).

Arrangement on gel:

DNA Ladder: FastRuler™ Middle Range, ready-to-use, 100-5000 bp Fermentas