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

From OpenWetWare

Jump to: navigation, search




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  

Contents

Competent Cells

  • New competent cells were made - we have a new stock in the -80[[:Category:{{{1}}}|{{{1}}}]] 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

LoxP PCR

  • With any luck this should be the final attempt
  • Ran along side the maxiprepped DNA
  • Loaded 10μL 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)
Personal tools