Team:TorontoMaRSDiscovery/22 August 2009


August 22, 2009

  1. Miniprepped overnights:
    • Tet1: Did 2 minipreps, one with normal protocol, one with low copy plasmid protocol
      • Normal protocol gave a higher yield
    • Tet with ampicillin: used low copy plasmid protocol, but cell pellet was much bigger therefore got the same yield as Tet1 normal protocol
  2. Digested the 2 samples with higher yields (500ng)
  3. Ran digests on gel, but still couldn't see ccdb gene
  4. Started to doubt restriction enzyme function, therefore redigested Tet H (1ug) and C plasmid (500ng)
  5. (1.5h digests for all digests today)
  6. Ran Tet H and C digests on a gel
    • A faint ~700bp band was observed in the Tet lane
    • More plasmid should be used for plasmid digests
  7. Started overnight ligation of 4+5 in Tet backbone