User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/07/28

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Objective

  • Transformation showed no colonies, so run a gel with ligation reaction from 7/26 and others.

Bench work

  1. Midiprep of pTXB1 and pTXB1.His out of DH10B (Promega PureYield™ Plasmid Midiprep system)
    • DNA quantitation of purified samples
  2. Run gel:
    • Lane 1: Ladder
    • Lane 2: ----
    • Lane 3: pTXB1 midiprep 7/28
    • Lane 4: pTXB1.His midiprep 7/28
    • Lane 5: pTXB1.His + BSA ligation 7/27
    • Lane 6: pTXB1.His (-)control ligation 7/27
    • Lane 7: pTXB1 +BSA ligation 7/27
    • Lane 8: pTXB1 (-)control 7/27
    • Lane 9: ----
    • Lane 10: pTXB1 w/ EcoRI + BamHI and phosphatase digest 7/26
    • Lane 11: pTXB1 w/ EcoRI + BamHI and phosphatase digest; ligase rxn 7/26
    • Lane 12: pTXB1 w/ EcoRI + BamHI 7/26
    • Lane 13: pTXB1 w/ EcoRI + BamHI; ligase rxn 7/26
    • Lane 14: ----
  3. Replated transformations from yesterday
  4. Re-ligated test ligations with newly aliquot'd ligase buffer. This due to concerns that it was not completely thawed, mixed, or otherwise homogenized before aliquot.

Results

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Gel

Analytical7.28labeled.jpg