• 800-930: Lu Chao: minipreps for 5xE7-term colonies and 7xlysis-term colonies. Prepare 2% gel.
• 1330-1400: Hung: digestion of the above plasmids with EcoRI and PstI for checking. Incubate for 2 hours.
• 1600-1815: Hung: run gel check:
  • 4 among 5 samples of E7-Term showed the exact bands that we're waiting for: 1 band at 2.2kb (E7-Term) and 1 band at 3kb (Term vector). Also, there's no band at 130bp, which means there's no uncut Terminator plasmid involved. The 5th sample of E7-Term has 1 band at 5kb (which should be E7-Term + Term Vector), and 1 weird band slightly below 2kb.
  • All 7 samples of Lysis-Term showed exact bands: 1 band slightly above 300bp (which should be Lysis-Term as Lysis is slightly less than 200bp and Term is 130bp); 1 band at 2kb (standard vector).
    

WELL! So we only have 2 more ligations to do: LacI-RBS with E7-Term and LsrA-RBS with Lysis-Term !

from left to right: 1kb ladder, 5xE7-Term, 100bp ladder, 7xLysis-Term