IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/11

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LacI + RBA-TesA ligation

  • Antarctic phosphotase treatment of LacI sample
Antarctic Phsophotase Treatment
  • Gel Extraction of RBS-TesA part: Sample was split evenly between 2 wells. Each well was collected separately and run through the standard procedure of the extraction kit (Zippy).
  • DNA Concentrations
  • LacI: 22.13 ng/μL
  • RBS-TesA: 17.985 ng/μL
  • ACC plasmid: 54.2 ng/μL
  • Ligation reaction
TesA+RBS insert with LacI+backbone
Reagent
LacI DNA 10.0
Antarctic Phosphotase 1.0
Phosphotase Buffer 2.0
H2O 7.0
37°C for 1 hour then 65°C for 5 min
Reagent 1 (μL) Control (μL)
TesA+RBS 2.0 0.0
LacI 2.0 2.0
Ligation Buffer 1.0 1.0
Ligase 1.0 1.0
H2O 4.0 6.0
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