User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/22

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Objective

Make media/buffers for growth and expression

Culture colony from glycerol stock-streaked plates from this weekend

Run DNA gel with pMXB10 2-stage QuikChange from Friday

Bench work

  1. Took 1 colony from each plate and placed into 3mL LB with 3μL 100mg/mL ampicillin (100μg/mL) and 15μL 40% w/v glucose (0.2% w/v)
    • Grew in 3mL culture for about 6 hours, shaking at 37°C
  2. Ran DNA gel (1% agarose) with pMXB10 2-stage QuikChange from previous week
    • Lane 1: Ladder mix, 10μL (10μL ladder mix + 2μL 6x loading buffer)
    • Lane 4: pMXB10 QC +His 8/19, 5μL sample + 1μL 6x loading buffer
    • Lane 7: pMXB10 QC Forward stage 1 8/19, 5μL sample + 1μL 6x loading buffer
    • Lane 10: pMXB10 QC Reverse stage 1 8/19, 5μL sample + 1μL 6x loading buffer

Results

GEL

DNAgel 8.22.2011 labeled.jpg