Devices demo
process
1. Put the consumptive material into the devices and use the holder to fix it in position 2. The holder rotates for bacteria activation 3. Put the sample on the elevator 4. Elevator move up and down 5. Suck the sample up by capillary phenomenon 6. The optical system work 7. The holder rotates for the nest sample detection 8. Finish detection movie demo
Shell
what is it for in our device?
Thermo Scientific™ Multiskan™ FC Filter-based Microplate Photometer Filter: 595 nm
Material
1 ml LUDOX mQH2O 96 well cell culture plate (clear with flat-bottom)
Method
1. Add 100 μl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette) 2. Add 100 μl of H2O into wells A2, B2, C2, D2 (or 1 mL H2O into cuvette) 3. Measure absorbance 600 nm of all samples in all standard measurement modes in instrument 4. Record the data in excel and Import data into Excel ( OD600 reference point tab ) Sheet_1 provided
Data result
Cell measure
Material
Competent cells ( Escherichia coli strain DH5α) LB (Luria Bertani) media Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 μg /mL) 50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light) Incubator at 37°C 1.5 ml eppendorf tubes for sample storage Ice bucket with ice Pipettes 96 well plate(cell culture 96 well plate、tissue culture testplate) Devices (from InterLab Measurement Kit): 1. Negative control(BBa_R0040) 2. Positive control(J23151+B0032+E0040+B0010+B0012) 3. Test Device 1: J23101+I13504 4. Test Device 2: J23106+I13504 5. Test Device 3: J23117+I13504 6. Test Device 4: J23101+BCD2+E0040+B0015 7. Test Device 5: J23106+BCD2+E0040+B0015 8. Test Device 6: J23117+BCD2+E0040+B0015
Method
1. Day 1 : Resuspended each plasmid in plate 7 and transform into Escherichia coli DH5α. (Transformation protocol is from iGEM) 2. Day 2 : Pick 2 colonies from each of plate and inoculate it on 5-10 mL LB medium +Chloramphenicol.Grow the cells overnight (16-18 hours) at 37°C and 170 rpm. 3. Day 3 : Set instrument to read OD600 (as OD calibration setting)and measure OD600 of the overnight cultures 4. Dilute the cultures to a target OD 600 of 0.02 in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube (covered with foil to block light). 5. Incubate the cultures at 37°C and 170 rpm. 6. Take 1000 μL samples of the cultures at 0, 2, 4, and 6 hours of incubation and place samples on ice 7. 4 replicates of 100 uL samples were taken from each culture at 0, 2, 4, and 6 hours of incubation and placed in a 96 well plate for OD and fluorescence measurements using the setup described above
Data result
