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Team:Warsaw/Calendar-Main/31 July 2008
From 2008.igem.org
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<center><h4>Checking if degradation of fusion with OmpA is a result of Top10 proteases' activity (lon, iompt)</center>Piotr:</h4> | <center><h4>Checking if degradation of fusion with OmpA is a result of Top10 proteases' activity (lon, iompt)</center>Piotr:</h4> | ||
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Inoculation of OmpA_omega_A_alpha and OmpA_A_alpha with and without inductor (0,5IPTG) in <i>E. coli</i> Rosetta strain. | Inoculation of OmpA_omega_A_alpha and OmpA_A_alpha with and without inductor (0,5IPTG) in <i>E. coli</i> Rosetta strain. | ||
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1. Optimization of PCR to obtain truncated fragment of protein A DNA Elongation time: 30s - Optimization of annealing temperature (gradient from 55°C to 75°C) 2. PCR to obtain truncated A protein DNA fragment Elongation time: 30s Annealing temperature: 60°C 20 cycles 3. Gel electrophoresis and isolation of 250 bp band.
Checking if degradation of fusion with OmpA is a result of Top10 proteases' activity (lon, iompt)Inoculation of OmpA_omega_A_alpha and OmpA_A_alpha with and without inductor (0,5IPTG) in E. coli Rosetta strain.
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