IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/17

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Ligation #3 Continued

  • Yesterdays streaked plates from the third ligation showed no growth, except for mCherry, which had maybe 15 very small colonies.
  • The mCherry that was poured out onto a new plate yesterday had a full lawn.
  • Also, the mCherry test of the competent cells grew very well. the 1.5 ml tube had been left in the incubator over night.
    • Considering this information, the remaining samples of the TesA/RBS ligation and the RBS backbone were poured onto new plates in the hopes that they too will show more growth.
    • Plates were left upstairs in the 32°C incubator, as the incubator downstairs doesn't hold a constant temp, and its airflow drys out the plates prematurely.
  • Seeds of the mCherry plate that grew were started.
    • Three seeds were made using 5 ml LB with chlor. antibiotic, and one colony. The pipette tip was dipped into the first and second and then scraped along the bottom of the third.