IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/06

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Entry title Checked plates from 8/5 Only + control on chloro and RBS-TesA on cholro had growth. Obtained acc plasmid (chloro resistance). Started plate and liquid colonies. Ethanol: At 8:30 AM, O.D. of cells was approx. 1.2 Resuspended in 30 ml PBS, adjusted O.D. to match at 1.0, resuspended in PBS again Began resting cell assay (9:15 AM): -Flask 1: 1 g/L pyruvate (.03 g) -Flask 2: 3 g/L pyruvate (.09 g)+ 1 g/L glucose (.03 g) -Flask 3: 3 g/L pyruvate (.09 g)(no glucose) Flasks were kept in 32°C shaker/incubator with 1 ml samples taken for Hrs. 1,2,3,6, and 8. (24 Hr. sample will be taken tomorrow) Made standards of ethanol and pyruvate to be run on the HPLC: Standards were first made of the max possible amount of each substance present in the system. (in 30 ml) These concentrations were then used as a stock to be diluted for subsequent concentrations Ethanol Standard Desired Concentration (mol/L) Water Added (ml) Pure Ethanol Added (ml) 1.13 28.021 1.979 .757 .33 .620 .38 .664 .336 .3 .735 .265 .2 .823 .177 .1 .912 .088 Pyruvate Standard Desired Concentration (g/L) Water Added (ml) Pyruvate Added (ml) 3.0 30 ml .09 g 2.0 .333 .667 1.0 .666 .333 .7 .767 .233 .5 .833 .167 .3 .900 .100