User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/03

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Bench work

  1. Re-ran QuikChange on pMXB10 using the 2 stage QuikChange method
    • (8 minutes each at elongation step x 17 repeats)
    • Stored at -20°C on completion
  2. Cut pKK223-3 plasmid (for a smaller-scale test ligation) with BamHI, which cuts this sample twice
    • 10μL 128ng/μL pKK223-3 from miniprep by Kay Wang 7/26/11
    • 5μL NEB3 10x + 5μL BamHI + 0.5μL BSA 100x + 29.5μL dH2O
    • After 2 hours @ 37°C, cleaned with Promega Wizard® DNA clean up system
  3. Replated yesterday's transformation
  4. Ligation of BamHI/EcoRI digest of pTXB1 from yesterday in addition to BamHI digest from today:
    • 10μL sample + 7μL dH2O + 1μL T4 DNA ligase
    • Two different buffers were used for each sample's ligase reactions -- an 'Old' aliquot and a 'New' one. 2μL were added to each respective reaction solution.
    • 16°C O/N

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