Difference between revisions of "Team:Bielefeld-CeBiTec/Notebook/CFPS"
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| − | + | <p> 04/22: Transformation of electrocompetent <i>E. coli</i> KRX with BBa_K1365020 (sfGFP (Bs)) | |
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| + | <p> 04/23: preparing glycerol stock of KRX with BBa_K1365020 | ||
| + | <p> 04/24-04/25: Mini-prep of BBa_K1365020 </p> | ||
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| − | <p>...</p> | + | <p>04/28: Transformation of electrocompetent <i>E. coli</i> KRX with BBa_I746909 (T7-promoter, RBS, sfGFP, double terminator). Before plating on LB(Cm), 50 µl of 200 g∙L<sup>-1</sup> Rhamnose-solution is added to the plate to induce T7-Polymerase. Inoculation of 10 ml preculture of simple KRX from plate. Overnight culturing, shaking vigorously at 220 rpm.</p> |
| + | <p>04/29: colonies of KRX with BBa_I746909 look greenish! 2.2 ml Overnight cultures from KRX are used to inoculate 100 ml for start OD<sub>600</sub>=0.144. Measuring of growth curves for a first estimation of appropriate moment for cell harvest (see image below) </p> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/e/e8/Bielefeld-CeBiTec_150429_growthcurveKRX.png" alt="Growth curves (duplicate) from <i> E. coli </i> KRX at 37 °C in 100 ml 2xTY+P-medium. 1 l shaking flasks without baffles were used. Shaking at 200 rpm" width="600" height="424"> | ||
| + | <p style="font-size:70%"> <b> Growth curves (duplicate) from <i> E. coli </i> KRX at 37 °C in 100 ml 2xTY+P-medium. 1 l shaking flasks without baffles were used. Shaking at 200 rpm. </b> </p> | ||
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Revision as of 10:20, 21 June 2015
Cell-free Protein Synthesis
04/22: Transformation of electrocompetent E. coli KRX with BBa_K1365020 (sfGFP (Bs))
04/23: preparing glycerol stock of KRX with BBa_K1365020
04/24-04/25: Mini-prep of BBa_K1365020
04/28: Transformation of electrocompetent E. coli KRX with BBa_I746909 (T7-promoter, RBS, sfGFP, double terminator). Before plating on LB(Cm), 50 µl of 200 g∙L-1 Rhamnose-solution is added to the plate to induce T7-Polymerase. Inoculation of 10 ml preculture of simple KRX from plate. Overnight culturing, shaking vigorously at 220 rpm.
04/29: colonies of KRX with BBa_I746909 look greenish! 2.2 ml Overnight cultures from KRX are used to inoculate 100 ml for start OD600=0.144. Measuring of growth curves for a first estimation of appropriate moment for cell harvest (see image below)
Growth curves (duplicate) from E. coli KRX at 37 °C in 100 ml 2xTY+P-medium. 1 l shaking flasks without baffles were used. Shaking at 200 rpm.