Solution Informationhelp
Enzyme: LuxS
inhibitor: BDBM65979
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Protocol Add 45 nL 10 mM sample / well to a sterile 384 well black clear bottom assay plate (Greiner microclear). Add 60 uL / well screening culture using a combi dispenser (Thermo) and read OD600 on Envision plate reader (Perkin-Elmer), incubate plate 9 h 30C in a humid incubator, read OD600 and luminescence on Envision plate reader. Quorum Sensing Reagents Screening Line JMH597 Vibrio harveyi (LuxN minus CQSSVH minus ) Overnight Culture 5 mL LM medium plus one colony JMH597, 30C 250 rpm Screening Culture Overnight Culture diluted to OD600 0.0005 in AB medium LM Medium (Rich medium for overnights and agar plates) 20 g NaCl (JT Baker, 3624-19) 10 g bactotryptone (BD, 211705) 5 g yeast extract (EMD, 1.03753) brought to 1 liter, 0.22 u sterile filter For plates, add 7.5 g agar (BD, 28130) / 500 ml LM autoclave 15 min, plate and flame AB Medium (Autoinducer Bioassay Medium) 100 ml 10X AB salt 2 g casamino acids (BD, 223050) 10 mL 1 M Phosphate Buffer pH 7 10 mL 0.1 M Arginine 100
 
 

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