IGEM:Wisconsin-Madison/2009/Notebook/Spring Training 2012/2012/03/07

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Plasmid extraction for training lab

I (EW) started 3x 10 mL liquid cultures yesterday evening (~7p) of K208000-pSB1AK3 and K634004-pSB1A2. NOTE: K208000 doesn't have an RBS, and will not work. TEACHABLE MOMENT >_<

  • Yishen Qiagen miniprepped one of each, and I did as well (One pair got mixed up and was thrown out).
  • Yishen's J23100-exaDE got two doses of N2.
  • All were eluted in 35 ul EB
  • Good yields, proper size bands.

Transforming E0240 for training lab

I (EW) want to try to put an RBS'd GFP in behind K634004 so that the lab can get some green fluorescence results. I'm taking E0240 from the 2009 spring distribution (2010 distro's was used and not freezer stocked or kept in clone box. Where the hell did 2011's go??), but since we want to use it tomorrow will be doing an hour outgrowth on non-selective media then inoculating LB+Amp100 (6 x 5 mL) and hoping for the best.

  • Transformed 1 uL of E0240 into a 2011 comp cell. Rest is in -20 labeled "2009 E0240"
    • Time constant: 3.20, outgrowth (5:15-6:15)