Team:UIUC-Illinois/Results/Scaffold

From 2012.igem.org

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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, 305 nM concentration of WT PUF-PIN, and 45 nM concentration of 6-2/7-2 PUF-PIN 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, 305 nM concentration of WT PUF-PIN, and 45 nM concentration of 6-2/7-2 PUF-PIN used.
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<img src="http://2012.igem.org/wiki/images/7/75/10.03.12_RNA_Scaffold_Assay.png"></center>
<img src="http://2012.igem.org/wiki/images/7/75/10.03.12_RNA_Scaffold_Assay.png"></center>
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<b>Fig. 7. </b>
<b>Fig. 7. </b>
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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. 3 mM MgCl2 ions used in all lanes. 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, 305 nM concentration of WT, 6-2/7-2 PUF-PIN in first four lanes, 152.5 nM concentration of WT, 6-2/7-2 PUF-PIN in last four lanes.  
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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. 3 mM MgCl2 ions used in all lanes. RNA samples were denatured for 3 min. at 95°C and then let to fold at 4°C for 5 min. Addition of 1 μL of annealing buffer (EDTA/Tris) before addition of protein. 30 min. incubation time with protein at 37°C and then addition of 2X 80% formamide/EDTA to stop the reaction. 66 nM concentration of RNA, 305 nM concentration of WT, 6-2/7-2 PUF-PIN in first four lanes, 152.5 nM concentration of WT, 6-2/7-2 PUF-PIN in last four lanes.  
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Latest revision as of 05:26, 26 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.

    Retrieved from "http://2012.igem.org/Team:UIUC-Illinois/Results/Scaffold"