IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/24

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Successful Ligation

  • Checked seeds from yesterday, all grew enough for miniprepping.
  • Ran a digest of the three colonies picked of the ligation, as well as the ethanol plasmid, to ensure that iGEM sent us the right part, and that the ligation was successful.
    • lane 7 was the only lane that showed a band that was the expected length for TesA+RBS. (This was colony #3 from plate #4)
    • Ethanol plasmid was as expected. (insert=3650, vector=2000)
Reagent Volume Lanes+Length:
1. 1 kb Ladder
3. TesA+RBS = 650
5. TesA+RBS = 650
7. TesA+RBS = 650
9. ETOH= 3650(ETOH backbone=2000)
DNA(plasmid) 3.0 μL
10X buffer 1.0
PSTI 1.0
EcoRI 1.0
dH2O 4.0
  10.0 μL --> 37°C/ ~35 min.