Team:Harvard/Results/Zinc Finger Binders

From 2011.igem.org

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(Color Blindness (CB) Bottom Hits)
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We did not have time to test all 6 targets as planned: we chose to focus on zinc finger binders to DNA tripplet TGG, which is found in the bottom color blindness gene.  
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We did not have time to test all 6 targets as planned: we chose to focus on zinc finger binders to DNA tripplet TGG, which is found in the color blindness gene bottom target.  
=Color Blindness (CB) Bottom Hits=
=Color Blindness (CB) Bottom Hits=
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We assembled the Color Blindness Bottom library, transformed it into the proper selection strain, and plated on incomplete media with varying concentrations of 3-AT.  We saw colonies form, with more on the less stringent plates (NM) and fewer colonies on the higher 3-AT concentrations. Several colonies were picked and tested under varying concentrations of 3-AT in plate reader overnight, and there was a clear difference in growth between the colonies and the negative control (CB bottom binding site with Zif268).  After sequencing, we identified xx novel zinc fingers!
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We assembled the Color Blindness Bottom library, transformed it into the proper selection strain, and plated on incomplete media with varying concentrations of 3-AT.  We saw colonies form, with more on the less stringent plates (NM) and fewer colonies on the higher 3-AT concentrations. Several colonies were picked and tested under varying concentrations of 3-AT in plate reader overnight, and there was a clear difference in growth between the colonies and the negative control (CB bottom binding site with Zif268).  After sequencing, we identified several novel zinc fingers!
[[File: HARVcbbot_1mM.png|center]]
[[File: HARVcbbot_1mM.png|center]]
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Revision as of 05:31, 28 September 2011

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We did not have time to test all 6 targets as planned: we chose to focus on zinc finger binders to DNA tripplet TGG, which is found in the color blindness gene bottom target.

Color Blindness (CB) Bottom Hits

We assembled the Color Blindness Bottom library, transformed it into the proper selection strain, and plated on incomplete media with varying concentrations of 3-AT. We saw colonies form, with more on the less stringent plates (NM) and fewer colonies on the higher 3-AT concentrations. Several colonies were picked and tested under varying concentrations of 3-AT in plate reader overnight, and there was a clear difference in growth between the colonies and the negative control (CB bottom binding site with Zif268). After sequencing, we identified several novel zinc fingers!

HARVcbbot 1mM.png