User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/11

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Objective

  • Ligate pKK223-3 BamHI digest
  • Check transformants

Bench work

  1. New BSA PCR primer designed. There was an error in BSA-SapI-3' where the sticky end sequence was inverted. New sequence is: 5'-TGG TTG GCT CTT CTG CAG GCT AAG GCT GT-3' (naming it BSA-SapI-Rev)
  2. Ligated re-cleaned pKK223-3 BamHI digest from yesterday
    • Tube 1: 10μL pKK223-3 with 2μL buffer, 7μL water, and 1μL T4 DNA ligase
    • Tube 2: 15μL pKK223-3 with 2μL buffer, 2μL water, and 1μL T4 DNA ligase
    • → 16°C O/N
  3. Replated transformants from yesterday


Results

  • TRANSFORMATION
    • One colony each on unclean and clean sample transformation plates
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