IGEM:Indian Institue of Technology Madras/2009/Notebook/PLASMID - Plasmid Locking Assembly for Sustaining Multiple Insert DNA/2009/10/16

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  • 06.15 - inoculated 4 colonies each of DH5a and Ca in 3 ml LB without antibiotics and with 25ug/ml of 3ml LB respectively (inoculted 4 of them so that atleast some of them will show OD >0.1 and <0.5 after 4 hours)
  • 10.15 - measured the OD of each of the 8 samples
  • DH5a-1 - 0.026
  • DH5a-2 - 0.215
  • DH5a-3 - 0.270
  • DH5a-4 - 0.120
  • CA-1 - 0.141
  • CA-1 - 0.186
  • CA-1 - 0.103
  • CA-1 - 0.034
  • out of these, DH5a-4 and CA-1 were selected for further inoculation of the main experimental broth
  • Using the calculation that was done yesterday (also since the measurements took about half hour, we assumed that the OD might have changed by about 0.01), 400ul aliquots of DH5a was inoculted into each of the 4 tubes (no ab, amp, chl, amp-chl) and 350ul aliquots of CA was inoculted into each of the 4 tubes at 11.30
  • 11.30 - samples were prepared for imaging. no cells were seen under the microscope. so wait for 3 hours
  • 12.30 - inoculated 4 colonies of RFP in 1C3 containing cells into 3ml LB broth with chl conc of 25ug/ul and 4 colonies of the co-transformants of C1A-R221 into 3ml broth with amp-chl conc of 25ug/ul each for OD measurements.
  • 2.30
                                 BOOM

the microscope got screwed and we have nothing to image now. wait till monday to get it repaired. an awesome time to have things no working

  • 4.30 - measured the OD of the inoculated colonies
  • The RFP inoculations did not show any growth
  • The cotransformed cells had an average growth of just less than 0.1