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

AU CHEM-570 Lab Prep

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Bench work 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 Re-cleaned pKK223-3 sample (ran on gel above) Transformed 5μL of BamHI-digested pKK223-3: cleaned sample, not cleaned sample, and dilute (20X) sample 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 The QuikChange samples of both plasmids do not appear. There is likely an error in technique or buffer/enzyme.