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Team:Wageningen UR/Notebook/Proj1/July
From 2011.igem.org
July 13
BioBrick parts were digested using different combinations of restriction enzymes, ligated and run on a gel.
July 18
Gel extracion of the gel run on July 13th was performed. Liquid cultures were made.
July 19
Yesterday's liquid cultures were miniprepped.
July 30
Inoculated 500 mL erlenmeyers with 100 ml LB + 100 ug/ul. E. coli was grown overnight on LB + amp plates and directly used as inoculum. All cultures were done in duplo. E. coli strains used were: ptet-GFP R0062-GFP F2621-GFP RBS-GFP
Cultures were put in a shake incubator at 30°C, 300 rpm. Temperature was set to 30 degrees as GFP is degraded at higher temperatures. The OD600s of every culture were measured 2 hours after inoculation and subsequently each hour, by taking 1 mL samples from the culture. the OD was measured with a Hitachi U1500 spectrophotometer. LB+amp was taken as blank. Fluorescence was measured in a 96-wells plate on a fluorescent plate reader (Molecular Devices, M2 (toxicology). First blank measurements were done with 96 wells plates containing 50 ul LB+amp. The output corresponding with the specific wells was saved and used for further calculations.
OD600 data presented below
| Sample | no. 1 (2 hours) | no. 2 (2 hours) | no. 1 (4 hours) | no. 2 (4 hours) | no. 1 (6 hours) | no. 2 (6 hours) |
|---|---|---|---|---|---|---|
| RBSGFP | min 0.001 | min 0.001 | 0.021 | 0.004 | 0 | 0 |
| R0062 | min 0.016 | min 0.011 | 0.001 | 0.020 | 0 | 0 |
| ptetGFP | min 0.006 | min 0.004 | 0.011 | min 0.003 | 0 | 0 |
| F2621GFP | min 0.014 | min 0.003 | 0.009 | 0.011 | 0 | 0 |
