Team:UIUC-Illinois/Results/Scaffold

From 2012.igem.org

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<b>Fig. 4.</b> Click on Fig. 4. to view a higher resolution image. In-Vitro Transcription with MEGAscript® T7 Kit (Invitrogen)
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<b>Fig. 4.>BCA analysis of WT & 6-2/7-2 PUF-PIN proteins to determine concentration (uM). 2 mg/mL BSA concentration used. Click on Fig. 4. to view it in a higher resolution.
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<b>Fig. 5.</b>In-Vitro Transcription with MEGAscript® T7 Kit (Invitrogen)
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<b>Fig.5</b>
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<b>Fig. 6.</b>
10 well 10% 1mm urea denaturing acrylamide gel, post 2μg/mL EtBr staining for 20 min., destaining for 20 min. 1X TBE buffer, 120V for 55 min. First 4 wells include 1 μL of 30 mM MnCl2 ions, last 4 wells include 1 μL of 30 mM MgCl2 ions. RNA samples were denatured for 3 min. at 95oC and then let to fold at 4oC for 5 min. Addition of 1 μL of annealing buffer (EDTA/Tris) before addition of protein. 30 min. incubation time with protein at 37oC and then addition of 2X 80% formamide/EDTA to stop the reaction. 66 nM concentration of RNA used.  
10 well 10% 1mm urea denaturing acrylamide gel, post 2μg/mL EtBr staining for 20 min., destaining for 20 min. 1X TBE buffer, 120V for 55 min. First 4 wells include 1 μL of 30 mM MnCl2 ions, last 4 wells include 1 μL of 30 mM MgCl2 ions. RNA samples were denatured for 3 min. at 95oC and then let to fold at 4oC for 5 min. Addition of 1 μL of annealing buffer (EDTA/Tris) before addition of protein. 30 min. incubation time with protein at 37oC and then addition of 2X 80% formamide/EDTA to stop the reaction. 66 nM concentration of RNA used.  

Revision as of 22:34, 3 October 2012

Header

Scaffold

RNA Scaffold

  • RNA Scaffold Results Overview
  • RNA Scaffold Data
  • PUF Tethering Data
  • Conclusion
  • RNA Scaffold Overview


    The results covered in this section are of the experiments overviewed in the RNA Scaffold Design section.