Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/17

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(Autocreate 2013/09/17 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
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==mm/dd/yyyy==
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* Insert content here...
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'''12PM'''
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Pulled RhlI out at 12pm. pH looks between 7 and 8 (paper strip)
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{| {{table}}
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|-
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|OD650||GFP||pH
 +
|-
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|0.726||16194||7-8
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|-
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|}
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<br>
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Spun 1mL of cells, resuspended in 100ul water, checked GFP: 158.  
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<br>
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Split cells and LB into four-2mL tubes<br>
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Spun at 12.0g for 3min<br>
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Transferred 1mL LB supernatant to new tubes
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:One-2mL tube for later induction (stored at -20)
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:Three-15mL conical tubes<br>
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Added 1mL ethyl acetate to LB<br>
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:Vortex for 15 seconds, sit for 2min
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::Repeat 5 times
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::Fifth time,let sit for 10 minutes
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Transfer 1mL of the top layer to 1.5mL tube
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Let sit for 15-30 seconds
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Transfer top layer to new 15mL conical tube
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Store at -20°C
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<br>
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<br>
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'''3PM'''
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Pulled Rhl2 out<br>
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{| {{table}}
 +
|-
 +
|OD650||GFP in LB||GFP in H20||pH
 +
|-
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|0.759||17964||81097||8-9
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|-
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|}
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<br>
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Repeated extraction procedure<br><br>
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'''6PM'''
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Pulled Rhl3 out<br>
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{| {{table}}
 +
|-
 +
|OD650||GFP in LB||GFP in H20||pH
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|-
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|0.689||18554||71065||8-9
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|-
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|}
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<br>
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Repeated the extraction procedure for Rhl3<br><br>
 +
 
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Tranformations into DH5αT and BL21<br>
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#J27 (F2620-RFP)
 +
#RhlI 8/31/13
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#EsaI sender A3
 +
#GFP-Lux rec 1

Revision as of 17:42, 14 October 2013

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mm/dd/yyyy

12PM Pulled RhlI out at 12pm. pH looks between 7 and 8 (paper strip)

OD650GFPpH
0.726161947-8


Spun 1mL of cells, resuspended in 100ul water, checked GFP: 158.
Split cells and LB into four-2mL tubes
Spun at 12.0g for 3min
Transferred 1mL LB supernatant to new tubes

One-2mL tube for later induction (stored at -20)
Three-15mL conical tubes

Added 1mL ethyl acetate to LB

Vortex for 15 seconds, sit for 2min
Repeat 5 times
Fifth time,let sit for 10 minutes

Transfer 1mL of the top layer to 1.5mL tube Let sit for 15-30 seconds Transfer top layer to new 15mL conical tube Store at -20°C

3PM Pulled Rhl2 out

OD650GFP in LBGFP in H20pH
0.75917964810978-9


Repeated extraction procedure

6PM Pulled Rhl3 out

OD650GFP in LBGFP in H20pH
0.68918554710658-9


Repeated the extraction procedure for Rhl3

Tranformations into DH5αT and BL21

  1. J27 (F2620-RFP)
  2. RhlI 8/31/13
  3. EsaI sender A3
  4. GFP-Lux rec 1



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