Difference between revisions of "Part:BBa K2145101:Experience"

 
 
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[[File:Splates105108.png]]<br>Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG106 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.
  
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__NOTOC__
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
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===Applications of BBa_K2145101===
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[[File:Rfp_fl105108.png]]
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[[File:Gfp_fl105108.png]]
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Above is TX-TL fluorescence data for this set of plasmids. Positive control is a GFP gene on a strong promoter and strong RBS (pBEST-OR2-OR1-Pr-UTR1-deGFP-T500, available on addgene). This plasmid did NOT express either RFP or GFP in TX-TL when added at 2nM (with dCas expression plasmid and gRNA expression plasmid). Addition of inducers to TX-TL reactions with these plasmids did not affect expression.
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Sequencing on this plasmid was low-quality and inconclusive.
  
 
===User Reviews===
 
===User Reviews===
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Latest revision as of 23:50, 30 October 2016

Splates105108.png
Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG106 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.


Rfp fl105108.png Gfp fl105108.png

Above is TX-TL fluorescence data for this set of plasmids. Positive control is a GFP gene on a strong promoter and strong RBS (pBEST-OR2-OR1-Pr-UTR1-deGFP-T500, available on addgene). This plasmid did NOT express either RFP or GFP in TX-TL when added at 2nM (with dCas expression plasmid and gRNA expression plasmid). Addition of inducers to TX-TL reactions with these plasmids did not affect expression.

Sequencing on this plasmid was low-quality and inconclusive.

User Reviews

UNIQe90f8bf69e7a4ad2-partinfo-00000000-QINU UNIQe90f8bf69e7a4ad2-partinfo-00000001-QINU