Part:BBa_K1416004:Experience

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UNIQ7d6365c38735f0f4-partinfo-00000000-QINU

No review score entered. Aachen 2016, Authors: C.Bonerath, A. Hoeltken, V.Czotscher

Summary As working with the fluorescence based screening system established by Team Austin, Texas, we created some data, which may be helpful to other users. Addionally we created a new part, to make the screening system available through the registry of standard biological parts. Documentation of the improvement Cultivation conditions with High Throughput measurement In order to evolve a new aminoacylation synthetase for DMNBS in E.coli, transforming a mutation library into competent cells the following order worked to get a maximum output and equal optical densities: Transform into (BL21 DE3 gold + pFRY) on M9 solid, growth: 2 days, 37°C Pick into M9 liquid: Masterplate, growth: 2 days at 30°C, 900rpm, shaking diameter: 50 mm Replicate into M9 liquid, growth 2 days at 30°C, 900 rpm, shaking diameter: 50 mm Replicate into M9 liquid Screening plate, growth 2 days at 30°C, 900 rpm, shaking diameter:50 mm For positive screening, supplementation at the beginning: Induction with 100 µM IPTG 2mM DMNBS as final concentration For negative screening, supplementation at the beginning: Induction with 100 µM IPTG Measurement: Wavelength normalized fluorescence spectrum of mRFP1 normalized fluorescence spectrum of sfGFP Measurement: Evaluation Links

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