| Solution Information | help | |
| Enzyme: | Alkanal monooxygenase alpha chain | |
| inhibitor: | BDBM65978 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Protocol Add 30 uL / well screening culture using a Combi dispenser (Thermo). Add compounds to be tested with pintool. Incubate 30 min 30C in a humid incubator. Add 30 uL / well AI-2 and read OD600 on Envision plate reader (Perkin-Elmer), incubate plate 5 h 30C in a humid incubator, read OD600 and luminescence on Envision plate reader. Quorum Sensing Reagents MM32 Vibrio harveyi (LuxS- ), AI-2 Overnight Culture 5 mL LM medium plus one colony MM32, 30C 250 rpm Screening Culture Overnight Culture diluted to OD600 0.020 in AB medium AI-2 DPD (4,5-dihidroxy-2,3-pentadione) diluted to 50 nM in AB medium DPD spontaneously reacts with borate in the medium to form AI-2 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) 1 | |
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