User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/02

AU CHEM-570 Lab Prep

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Objective No growth on yesterday's plates → replate remainder this afternoon DpnI digest of +His/(-)control QuikChange of pMXB10 from yesterday; subsequently transform these samples Run gel with QC samples Double-digest of pTXB1 with BamHI and EcoRI (retrial with higher [DNA]) Bench work Ran 1.2% agarose gel with QC reactions Lane 1: Ladder Lane 3: pMXB10 QC +His from yesterday Lane 5: pMXB10 QC (-) yesterday Added 1μL of DpnI to each of the QC reactions from yesterday Double-digest of pTXB1 with BamHI and EcoRI NEB suggests NEBuffer EcoRI for this double-digest, but NEBuffer 3 was used instead. 50μL volume: 10μL 200μg/mL pTXB1, 5μL NEB 3 (10X), 5μL EcoRI (2kU/mL), 5μL BamHI (20kU/mL), 24.5μL dH2O, 0.5μL BSA (100X) 2 hours @ 37°C Results Gel