Virginia Commonwealth/24 June 2009

From 2009.igem.org

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Contents

Wednesday 24 June 2009

Results

  • Gel Electrophoresis was run on pSB3K3 only
  • No DNA was visible

2009-06-24 19digest02(psb3k3only).jpg

Trentay 19:16, 5 August 2009 (UTC)


Tasks

  • Miniprep needs to be done on regrown backbone stock.
  • Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240).
  • Run a gel electrophoresis on the digested PSB3K3.

Wetlab

  • The promoter DNA from the mini-preps run by Afton on the 12th were Concentrated using the "Ethanol precipitation of nucleic acids" protocol on OpenWetWare.(Detailed notes can be found on page 10 of the iGEM notebook.)
    • Note: A spec needs to be run on the concentrated DNA to obtain the current concentrations and volumes.
      • This should be held off until the backbone problems are rectified as the DNA may degrade over time.
  • Digestion was done on the PSB3K3 using the enzymes ECO RI-HF and PstI, in the respective order.
    • Problems:
      • The 80 deg. Celsius bath was not ready during the incubation so the cells were kept in the incubator longer than 15 minutes.
  • Ligation was performed along with transformation and is growing up overnight. The results will be listed above in the morning.
    • Problems:
      • Maria was distracted while transforming and used 5 increments of 200 uL of the SOC because the 1000 uL pipet was not ready. The delay may have caused decreased efficiency (although it was not more than 1 minute before the 1st 200uL).
  • Gel Electrophoresis was run, however no PSB3K3 was found on the plate. (details on page 12 of the iGEM notebook)
  • PSB3K3 was re-plated using the plasmid from the Biobrick well stock. Colonies should be picked tomorrow afternoon.

(All data is on pages 10-12 in the iGEM lab notebook for more detail.)

MandM 02:22, 25 June 2009 (UTC)