IGEM:IMPERIAL/2006/LabCalendar/2006-8-30

 TO DO: *Make Competent Cells *Electroporate J37015, J37024 (from ligations) after having made competent cells *Miniprep J37024, J37022 and J37025 (to check whether ligations were successful) *Religation of J37018 ? *Freeze 4G (S01656), J37020 (&J37018?) *Testing (T9002, J37016 & J37020) *PCR

Competent Cells

 * New competent cells were made - we have a new stock in the -80 freezer again

Measuring the growth rate of the prey cells

 * JohnChattaway 04:50, 30 August 2006 (EDT)
 * We need to do this to work out the growth rate of the prey to give us an idea of the amounts of media we need for the chemostat.IGEM:IMPERIAL/2006/The Talk Page
 * Cultured fresh cells at 9.30
 * At 11.30 will re-culture to OD of 0.1 then test OD every 30mins untill OD 0.9 is reached

Measuring AHL production overnight

 * JohnChattaway 04:50, 30 August 2006 (EDT)
 * I will repeat tom's experiment with the frozen Prey Cells

Testing T9002, J37016 and J37020

 * Took 16mL cultures out of shaker after 2h at 11:20
 * OD measurements:
 * T9002 0.186
 * J37016 0.218
 * J37020 0.160
 * Dilute down to OD 0.1 into 25mL
 * Put 5mL tubes with AHL in shaker at 12:00 - to be taken out at 16:00
 * Tubes were taken out at 16:25
 * Fluorescence reading of the 3 plates (17:50 - 19: 10)

Ligations

 * Part J37024 glass milk purified and electroporated.

Minipreps of J37025, J37022, J37018

 * See [[image:Miniprep_30-08.pdf|Miniprep_30-08.pdf|here]] for photo of gel
 * Comment: The part outlined in the photo as J37020 is actually J37022 !!

LoxP PCR

 * With any luck this should be the final attempt
 * Ran along side the maxiprepped DNA
 * Loaded 10 of each PCR reaction into the gel (double the amount of the previous attempts)

Freezing Cells

 * Froze S01656 and J37020 (the right ones this time - old ones have been thrown out)

Culturing

 * J37024 (from both plates for miniprep)
 * T9002/J37016/J37020/J37015/J37015RS/S01656 (for testing)