Virginia Commonwealth/10 July 2009

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Contents

Friday 10 July 2009

Results

Afton and Maria

  • Overnight culture was successful

Trentay 13:13, 10 July 2009 (UTC)

Craig and Clay

  • The overnight culture of the three parts related to limonene synthesis were unsuccessful. Possible reasons include bad parts, errors in electrotransformation procedure, and incorrect resistance.

Tasks

Afton and Maria

  • Make stocks of parts J23106 and J06702
  • Miniprep of parts J23106 and J06702
  • Digestion of parts J23106 and J06702
  • Electrophoresis test of parts J23106 and J06702
  • May Ligate parts with backbones pSB1C3 and pSB4C5 and store DNA
  • Write out complete plan for experimentation
    • Make plans to MaxiPrep
    • Write out a plan to organize ideas

Trentay 13:13, 10 July 2009 (UTC)

Craig and Clay

  • BBa_I742111, BBa_K118024, and BBa_K118025 need to be plated again.
  • If the new plates also show no growth, electrotransformation of the parts will be performed again.
  • If this proves to be unsuccessful, the cells from this second electrotransformation will also be plated a second time. The cells will also be plated on non-resistant media.
  • If growth is again unsuccessful, the cells grown on the non-resistant media will be streaked on kanamycin-resistant and chloramphenicol-resistant plates.

Wetlab

Afton and Maria

  • Dilution of overnight culture
    • 2 mL of overnight culture was added to 4 mL of LB broth (6 mL)
      • 2 dilutions were done for each test tube
    • Cells not used in the MiniPrep will be stored in the -80 degree Celsius freezer
  • 500mL of 30 percent Glycerol stock was made
  • Cell stocks were made
    • 10 vials of J06702 were put in the -80 degree Celsius freezer
    • 5 vials of J23106 were put in the -80 degree Celsius freezer
  • MiniPrep was done on parts: J23106 and J6702
    • Amount of DNA was calculated with a Spectrophotometer
  • Digestion was performed on MiniPrepped parts
    • There are two digested J06702 PCR tubes
    • Digests will be frozen until Monday
      • Electrophoresis test will be held off until then
      • Ligation as well

Trentay 20:35, 10 July 2009 (UTC)

Craig and Clay

  • The electrotransformed parts were plated again due to unsuccessful growth. Fifty microliters was used instead of 35 microliters.
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