Virginia Commonwealth/24 June 2009

Results

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

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)