Haynes Lab:Notebook/Engineering PC-TFs/2012/10/26

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Summary

Assembly # Concentration
1 24.41 ng/μL
2 17.025 ng/μL
3 42.325 ng/μL
4 16.706 ng/μL
5 45.015 ng/μL











  • Gibson Assembly

Assembly 1 contains PCR rxns 1,2, and 3. Assembly 2 contains PCR rxns 4,5.

1 2
Gibson Rxn mix 15 μL 15 μL
DNA fragments 5 μL 5 μL







DNA fragments 1 2
#1 1.0 μL
#2 1.0 μL
#3 1.0 μL
#4 1.0 μL
#5 1.0 μL
Vector 1.0 μL 1.0 μL
dH2O 1.0 μL 2.0 μL
Total 5.0 μL 5.0 μL

















  • Thaw DHSα Turbo cells (50 μL per transformation)
  • Add 20 μL of Gibson reaction to 50μL DHSα Turbo.
    • Assembly 1
    • Assembly 2
    • Negative Control (DH2O)
  • Incubate on ice for five minutes.
  • Plate 70 μL transformation on warm Amp plates.




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