Team:UIUC-Illinois/Results/Scaffold

From 2012.igem.org

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<br/><b>Fig. 1.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight for WT PUF-PIN and 6-2/7-2 PUF-PIN. 2 mM IPTG induction for 2 hours. 200 mL cultures incubated at 37oC till 1 nm optical density after inoculation with 2 mL of overnight for WT PUF-αGFP cultures. 2 mM IPTG induction for various hours.
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<br/><b>Fig. 1.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight for WT PUF-PIN and 6-2/7-2 PUF-PIN. 2 mM IPTG induction for 2 hours. 200 mL cultures incubated at 37oC till 1 nm optical density after inoculation with 2 mL of overnight for WT PUF-αGFP cultures. 2 mM IPTG induction for various hours. SDS-PAGE 6% native gel stained with Coomassie Brilliant Blue for 1 hr. and destained with H20 for 1 hr.
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<img src="http://2012.igem.org/wiki/images/4/41/WT_PUF-PIN_Protein_Purification.png" height=100% width=100%></center>
<img src="http://2012.igem.org/wiki/images/4/41/WT_PUF-PIN_Protein_Purification.png" height=100% width=100%></center>
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<br/><b>Fig. 2.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight. 2 mM IPTG induction for 2 hours. His-Tag Ni-NTA purification, centrifuged with Millipore 30kDa cutoff ultracentrifuge tubes.  
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<br/><b>Fig. 2.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight. 2 mM IPTG induction for 2 hours. His-Tag Ni-NTA purification, centrifuged with Millipore 30kDa cutoff ultracentrifuge tubes. SDS-PAGE 6% native gel stained with Coomassie Brilliant Blue for 1 hr. and destained with H20 for 1 hr.
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<img src="http://2012.igem.org/wiki/images/6/6f/6-2%2C7-2_PUF-PIN_Protein_Purification.png" height=100% width=100%></center>
<img src="http://2012.igem.org/wiki/images/6/6f/6-2%2C7-2_PUF-PIN_Protein_Purification.png" height=100% width=100%></center>
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<br/><b>Fig. 3.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight. 2 mM IPTG induction for 2 hours. His-Tag Ni-NTA purification, centrifuged with Millipore 30kDa cutoff ultracentrifuge tubes.  
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<br/><b>Fig. 3.</b> 1 L culture incubated at 37oC till 0.5 nm optical density after inoculation with 5 mL of overnight. 2 mM IPTG induction for 2 hours. His-Tag Ni-NTA purification, centrifuged with Millipore 30kDa cutoff ultracentrifuge tubes. SDS-PAGE 6% native gel stained with Coomassie Brilliant Blue for 1 hr. and destained with H20 for 1 hr.
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<b>Fig. 4. </b>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. 4. </b>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. SDS-PAGE 6% native gel stained with Coomassie Brilliant Blue for 1 hr. and destained with H20 for 1 hr.
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<b>Fig. 5. </b>In-Vitro Transcription with MEGAscript® T7 Kit (Invitrogen)
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<b>Fig. 5. </b>In-Vitro Transcription with MEGAscript® T7 Kit (Invitrogen)  
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Revision as of 03:22, 4 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"