IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/11: Difference between revisions

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* Antarctic phosphotase treatment of LacI sample
* Antarctic phosphotase treatment of LacI sample
: Antarctic Phsophotase Treatment
: Antarctic Phsophotase Treatment
{| {{table}} border="1" align=center <!-- Digestion table -->
{| {{table}} border="1" align=center  
|- valign="top"
|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Reagent
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*Ligation reaction
*Ligation reaction
: TesA+RBS insert with LacI+backbone
: TesA+RBS insert with LacI+backbone
{| {{table}} border="1" align=center <!-- Digestion table -->
{| {{table}} border="1" align=center  
|- valign="top"
|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Reagent

Revision as of 15:18, 11 August 2014

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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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