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

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2013/09/17 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
Line 7: Line 7:
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==mm/dd/yyyy==
==mm/dd/yyyy==
* Insert content here...
'''12PM'''
Pulled RhlI out at 12pm. pH looks between 7 and 8 (paper strip)
{| {{table}}
|-
|OD650||GFP||pH
|-
|0.726||16194||7-8
|-
|}
<br>
Spun 1mL of cells, resuspended in 100ul water, checked GFP: 158.  
<br>
Split cells and LB into four-2mL tubes<br>
Spun at 12.0g for 3min<br>
Transferred 1mL LB supernatant to new tubes
:One-2mL tube for later induction (stored at -20)
:Three-15mL conical tubes<br>
Added 1mL ethyl acetate to LB<br>
: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
<br>
<br>
'''3PM'''
Pulled Rhl2 out<br>
{| {{table}}
|-
|OD650||GFP in LB||GFP in H20||pH
|-
|0.759||17964||81097||8-9
|-
|}
<br>
Repeated extraction procedure<br><br>
'''6PM'''
Pulled Rhl3 out<br>
{| {{table}}
|-
|OD650||GFP in LB||GFP in H20||pH
|-
|0.689||18554||71065||8-9
|-
|}
<br>
Repeated the extraction procedure for Rhl3<br><br>
 
Tranformations into DH5αT and BL21<br>
#J27 (F2620-RFP)
#RhlI 8/31/13
#EsaI sender A3
#GFP-Lux rec 1




Revision as of 14:42, 14 October 2013

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

mm/dd/yyyy

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

OD650 GFP pH
0.726 16194 7-8


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

OD650 GFP in LB GFP in H20 pH
0.759 17964 81097 8-9


Repeated extraction procedure

6PM Pulled Rhl3 out

OD650 GFP in LB GFP in H20 pH
0.689 18554 71065 8-9


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



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs/2013/09/17&oldid=738236"