User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/10

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Bench work

  1. DNA gel with QuikChange samples from yesterday
    • Lane 1: Ladder
    • Lane 2: ----
    • Lane 3: pKK223-3 clean-up, BamHI from yesterday (Could not load)
    • Lane 4: ----
    • Lane 5: pTXB1 QuikChange +His 8/9
    • Lane 6: pTXB1 forward, stage 1 of QC +His 8/9
    • Lane 7: pTXB1 reverse, stage 1 of QC +His 8/9
    • Lane 8: ----
    • Lane 9: pMXB10 QuikChange +His 8/9
    • Lane 10: pMXB10 forward, stage 1 of QC +His 8/9
    • Lane 11: pMXB10 reverse, stage 1 of QC +His 8/9
  2. Re-cleaned pKK223-3 sample (ran on gel above)
  3. Transformed 5μL of BamHI-digested pKK223-3: cleaned sample, not cleaned sample, and dilute (20X) sample
  4. Test QC with pTXB1:
    • 2-stage QuikChange with two different Pfu turbo buffers
    • 5 cycles in stage 1, 17 in stage 2; 7 minute elongation


Results

  • TRANSFORMATION
    • One colony on non-cleaned pKK223-3 transformation plate, none on others

Gel

Image: gel8.10.2011.jpg

  • The QuikChange samples of both plasmids do not appear. There is likely an error in technique or buffer/enzyme.
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