Team:Hamburg/InterLab
InterLab Study
Interlab study
To allow the comparison of consistency in measurement between all participating teams all over the world the interlab study takes place as a benchmark. It is organized by the iGEM's measurement committee and serves the idea to improve measurement techniques in the field of synthetic biology. Therefore, all participating teams will perform the same biochemical experiments, the measurement of GFP and OD600 of the DH5α E. coli cells of samples provides by the iGEM's headquarter.
Description of the experiment:
The following devices were available used for the experiments:
- Positive control
- Negative control
- Test Device 1: J23101 + I13504
- Test Device 2: J23106 + I13504
- Test Device 3: J23117 + I13504
- Test Device 4: J23101.BCD2.E0040.B0015
- Test Device 5: J23106.BCD2.E0040.B0015
- Test Device 6: J23117.BCD2.E0040.B0015
The samples were transformed into the DH5 α E. coli strain by the following transformation protocol:
- Thaw cells on ice
- Add DNA and mix
- Incubate on ice for 1 h
- Heat-shock 1 min 4 °C
- Incubate 4 min on ice
- Add 850 µl 37 °C Luria-Bertani media (LB media) to cells
- Incubate 1 h 37 °C 900 rpm
- Centrifuge 4500 rpm 2 min
- Remove 850 µl liquid
- Resuspend cells in remaining liquid
- Grease cell suspension on plates with cAMP resistance
- Incubate overnight 37 °C
The cells were applied to plates with a chloramphenicol (cAMP) antibiotic with a concentration of 25 µg/ml and incubated over night at 37 °C.
After 16 - 18 hours of incubation two positive colonies for each transformation were transferred to 5 ml liquid media with a chloramphenicol antibiotic with a concentration of 25 µg/ml. The cells were incubated over night at 37 °C at 220 rpm.
The densities of the cultures were measured at OD600 The Tecan Genios Microplate reader was used for the measurement. The samples were diluted to an OD600 = 0.02 in 12 ml LB media with chloramphenicol concentration of 25 µg/ml. The samples were incubated at 37 °C and 220 rpm. Samples of 500 µl were taken after 0, 2, 4 and 6 hours and stored on ice for further experiments. 100 µl of each sample was given into a 96-well plate as in the following pattern:
To allow the comparison of consistency in measurement between all participating teams all over the world the interlab study takes place as a benchmark. It is organized by the iGEM's measurement committee and serves the idea to improve measurement techniques in the field of synthetic biology. Therefore, all participating teams will perform the same biochemical experiments, the measurement of GFP and OD600 of the DH5α E. coli cells of samples provides by the iGEM's headquarter.
Description of the experiment:
The following devices were available used for the experiments:
- Positive control
- Negative control
- Test Device 1: J23101 + I13504
- Test Device 2: J23106 + I13504
- Test Device 3: J23117 + I13504
- Test Device 4: J23101.BCD2.E0040.B0015
- Test Device 5: J23106.BCD2.E0040.B0015
- Test Device 6: J23117.BCD2.E0040.B0015
The samples were transformed into the DH5 α E. coli strain by the following transformation protocol:
- Thaw cells on ice
- Add DNA and mix
- Incubate on ice for 1 h
- Heat-shock 1 min 4 °C
- Incubate 4 min on ice
- Add 850 µl 37 °C Luria-Bertani media (LB media) to cells
- Incubate 1 h 37 °C 900 rpm
- Centrifuge 4500 rpm 2 min
- Remove 850 µl liquid
- Resuspend cells in remaining liquid
- Grease cell suspension on plates with cAMP resistance
- Incubate overnight 37 °C
The cells were applied to plates with a chloramphenicol (cAMP) antibiotic with a concentration of 25 µg/ml and incubated over night at 37 °C.
After 16 - 18 hours of incubation two positive colonies for each transformation were transferred to 5 ml liquid media with a chloramphenicol antibiotic with a concentration of 25 µg/ml. The cells were incubated over night at 37 °C at 220 rpm.
The densities of the cultures were measured at OD600 The Tecan Genios Microplate reader was used for the measurement. The samples were diluted to an OD600 = 0.02 in 12 ml LB media with chloramphenicol concentration of 25 µg/ml. The samples were incubated at 37 °C and 220 rpm. Samples of 500 µl were taken after 0, 2, 4 and 6 hours and stored on ice for further experiments. 100 µl of each sample was given into a 96-well plate as in the following pattern:
