IGEM:IMPERIAL/2006/LabCalendar/2006-9-2

• IGEM:IMPERIAL/2006/LabCalendar/2006-9-2 Events
• Day in the lab
• Project Progress

TO DO:
*Testing of T9002, J37016, J37020, S01656, J37015, J37015RS
*Culture up J37022 and J37025 from plates in incubator for mini & maxi on Sunday
*Move 3 plates (2 Rosetta and 1 BL21) into cold room

Testing of T9002, J37016, J37020

• Reported PROBLEM: After the 2hrs shake of the 16mL cultures, the OD of the T9002 assay was still at about 0.1. So all of the present culture was diluted further into 25mL.
• 5mL tubes put into shaker for 4hrs at 13:15
• Taken out of shaker at 17:20

OBSERVATION: Both cultures of J37016 & J37020 looked very transparent, whereas T9002 was very "milky" (high OD).
Was the OD read correctly when diluting into 25mL ?

• Pipetting into 96 well plate

PROBLEM: There was not enough cell culture in some of the 5mL tubes to fill all 8 repeats of the wells. There was only about 1.5mL culture in some of the tubes. In others, there was about 1.8mL. Why?

• Fluorescence reading in Biochem (18:00)
• Results:
  • T9002 shows a slight increase of flurescence with AHL increase (however, it is a line, not reaching saturation)
  • J37016 do show the expected response, however, a sudden trough can be seen in at [AHL]=1nM
  • J37020 no change in fluorescence, apart from a trough at [AHL]=7nM
  • See results page for data

Culturing J37022 and J37025

• Cultured J37022 and J37025 from plate for mini and maxiprep tomorrow