Difference between revisions of "Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/05/22"

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(Autocreate 2014/05/22 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
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PCR clean up of EsaI PCR and mCh Sender Intermediate for golden gate. Forgot to do DpnI digest of mCh (don't need to for EsaI because the PCR template is kan resistance).  
  
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Sample'''
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| align="center" style="background:#f0f0f0;"|'''260'''
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| align="center" style="background:#f0f0f0;"|'''280'''
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| align="center" style="background:#f0f0f0;"|'''260/280'''
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| align="center" style="background:#f0f0f0;"|'''ng/µL'''
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|-
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| mCh Sender int gg||0.055||0.031||1.774||55.337
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|-
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| EsaI||0.046||0.026||1.776||45.692
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|}
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<br><br>
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Going to run golden gate anyway. Backbone plate should have the same number of PCR template transformants as any of the ligation plates.
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Reaction'''
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| align="center" style="background:#f0f0f0;"|'''Rec Bb'''
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| align="center" style="background:#f0f0f0;"|'''LuxI'''
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| align="center" style="background:#f0f0f0;"|'''BsmBI'''
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| align="center" style="background:#f0f0f0;"|'''T4 Ligase'''
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| align="center" style="background:#f0f0f0;"|'''T4 Ligase buffer'''
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| align="center" style="background:#f0f0f0;"|'''H2O'''
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|-
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| Bb ctrl||0.77||0||0.5||1.25||2||15.48
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|-
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| RhlI||0.77||3.2||0.5||1.25||2||12.28
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|-
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| LasI||0.77||2.8||0.5||1.25||2||12.68
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|-
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| EsaI||0.77||1.9||0.5||1.25||2||13.58
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|}
  
 
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Revision as of 11:05, 22 May 2014

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mm/dd/yyyy

PCR clean up of EsaI PCR and mCh Sender Intermediate for golden gate. Forgot to do DpnI digest of mCh (don't need to for EsaI because the PCR template is kan resistance).

Sample 260 280 260/280 ng/µL
mCh Sender int gg 0.055 0.031 1.774 55.337
EsaI 0.046 0.026 1.776 45.692



Going to run golden gate anyway. Backbone plate should have the same number of PCR template transformants as any of the ligation plates.

Reaction Rec Bb LuxI BsmBI T4 Ligase T4 Ligase buffer H2O
Bb ctrl 0.77 0 0.5 1.25 2 15.48
RhlI 0.77 3.2 0.5 1.25 2 12.28
LasI 0.77 2.8 0.5 1.25 2 12.68
EsaI 0.77 1.9 0.5 1.25 2 13.58