Objective
- Transformation showed no colonies, so run a gel with ligation reaction from 7/26 and others.
Bench work
- Midiprep of pTXB1 and pTXB1.His out of DH10B (Promega PureYield™ Plasmid Midiprep system)
- DNA quantitation of purified samples
- 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: ----
- Replated transformations from yesterday
- 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
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AU CHEM-570 Lab Prep
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