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TUDelft/30 September 2008
From 2008.igem.org
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September 30th
3A Assembly Thermoparts
K115022, K115036 (from a frozen pellet [1]) and K115027 were miniprepped.
After plasmids were obtained, restrictions have been started for K115022, K115024, K115025 and K115027 with XbaI and PstI. R0080 was cut with EcoRI and SpeI. pSB1AT3 was cut with EcoRI and PstI.
After restrictions were completed, ligations were started o/n for K115030, K115032, K115033 and K115035.
Protein Assay
We've made a new attempt at a protein assay, because lysis buffer showed to interfere with the measurements in the last assay. To try to compensate we've made a dilution series of BSA in 1000x diluted lysis buffer to make a new calibration line and measure again. Samples of lysed cultures of different temperatures were also diluted 1000x. The new measurements indicated that some samples had "negative" protein content indicating that lysis by Promega lysis buffer is a bad option for our current assay. We will have to look into non-interfering protein assays or optimize lysis by sonication.
