Haynes Lab:Notebook/Engineering PC-TFs/2013/03/27

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3/27/2013

  • 1. Kozak,V0120 + BL01
  • 2. Kozak,V0120 + BL05
  • 3. Kozak,V0120 + BL09
  • 4. Kozak,V0120 + BL12
1 2 3 4 5
DNA Insert 1.4 1.8 2.0 2.5 --
DNA Vector (Kozak) 0.6 0.6 0.6 0.6 0.6
T4 Ligase 1 1 1 1 1
Lign Buffer (2x) 5 5 5 5 5
dH2O 2.0 1.6 1.4 .9 3.7
10 µL 10 µL 10 µL 10 µL 10 µL
  • In a .5 mL tube, pipette the ligation buffer first.
  • Then water, DNA insert, vector, and T4 ligase.
  • Once completed, allow them to incubate for 10 minutes.
  • Meanwhile, get ice bucket and allow competent E. coli cells to thaw for five minutes.
  • Then follow transformation process:



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