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<h5>Cell Measurement Procedure</h5> | <h5>Cell Measurement Procedure</h5> | ||
Day 1: transform Escherichia coli DH5α with these following plasmids: | Day 1: transform Escherichia coli DH5α with these following plasmids: | ||
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● Positive control | ● Positive control | ||
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● Negative control | ● Negative control | ||
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● Test Device 1: J23101+I13504 | ● Test Device 1: J23101+I13504 | ||
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● Test Device 2: J23106+I13504 | ● Test Device 2: J23106+I13504 | ||
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● Test Device 3: J23117+I13504 | ● Test Device 3: J23117+I13504 | ||
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● Test Device 4: J23101.BCD2.E0040.B0015 | ● Test Device 4: J23101.BCD2.E0040.B0015 | ||
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● Test Device 5: J23106.BCD2.E0040.B0015 | ● Test Device 5: J23106.BCD2.E0040.B0015 | ||
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● Test Device 6: J23117.BCD2.E0040.B0015 | ● Test Device 6: J23117.BCD2.E0040.B0015 | ||
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Day 2: Picked 2 colonies from each of plate and inoculate it on 5-10 mL LB medium + Chloramphenicol. | Day 2: Picked 2 colonies from each of plate and inoculate it on 5-10 mL LB medium + Chloramphenicol. | ||
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Grow the cells overnight (16-18 hours) at 37°C and 220 rpm. | Grow the cells overnight (16-18 hours) at 37°C and 220 rpm. | ||
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Day 3: Cell growth, sampling, and assay | Day 3: Cell growth, sampling, and assay | ||
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◻ Set instrument to read OD600 (as OD calibration setting) | ◻ Set instrument to read OD600 (as OD calibration setting) | ||
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◻ Measured OD600 of the overnight cultures | ◻ Measured OD600 of the overnight cultures | ||
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◻ Recorded data | ◻ Recorded data | ||
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◻ Imported data into Excel (Dilution Calculation) Sheet_1 provided | ◻ Imported data into Excel (Dilution Calculation) Sheet_1 provided | ||
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◻ Dilute the cultures to a target OD600 of 0.02 (see the volume of preloading culture and | ◻ Dilute the cultures to a target OD600 of 0.02 (see the volume of preloading culture and | ||
media in Excel (Dilution Calculation) Sheet_1) in 15 ml LB medium + Chloramphenicol | media in Excel (Dilution Calculation) Sheet_1) in 15 ml LB medium + Chloramphenicol | ||
in 50 mL falcon tube (amber, or covered with foil to block light). | in 50 mL falcon tube (amber, or covered with foil to block light). | ||
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◻ Incubate the cultures at 37°C and 220 rpm. | ◻ Incubate the cultures at 37°C and 220 rpm. | ||
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◻ Take 1 mL samples of the cultures at 0, 2, 4, and 6 hours of incubation. (At each time | ◻ Take 1 mL samples of the cultures at 0, 2, 4, and 6 hours of incubation. (At each time | ||
point, you will take a sample from each of the 8 devices, two colonies per device, for a | point, you will take a sample from each of the 8 devices, two colonies per device, for a | ||
total of 16 samples per time point) | total of 16 samples per time point) | ||
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◻ Place samples on ice. | ◻ Place samples on ice. | ||
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◻ At the end of sampling point you need to measure your samples (OD and Fl | ◻ At the end of sampling point you need to measure your samples (OD and Fl | ||
measurement), see the below for details. | measurement), see the below for details. | ||
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◻ Record data in your notebook | ◻ Record data in your notebook | ||
<br/> | <br/> |
Revision as of 22:01, 29 September 2017
Florida_Atlantic