Team:Wageningen UR/Notebook/Proj1/July

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 with the Fermentas Gel Extraction kit. Liquid cultures were made, 10 mL LB + antibiotics in a 50 mL Greiner and grown o/n.

July 19

Yesterday's liquid cultures were miniprepped with the Fermentas miniprep kit.

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

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