Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/02/26: Difference between revisions

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(Autocreate 2014/02/26 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
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==mm/dd/yyyy==
==02/26/2014==
* Insert content here...
Spun overnight cultures of GFP-Senders at 13.1g for 10 minutes in 2mL tubes in benchtop centrifuge. Filtered using Fuqing's 0.45μm sterile filters and syringes. Made 5mL of 1% LuxI media. 5mL of LB+amp and 50ul LuxI sender.
 
<br>
Set up microfluidics with David Menn. LuxI inducing LuxR-GFP at 1%.
<br>
Made dilutions of filtered sender media. Wanted to make sure each concentration has the same ratio of spent media to fresh media. Made 1800ul. 300ul for 3 replicates of each dilution. 1500ul for flow cytometry. Can take some off the top at multiple time points.
<br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''%media'''
| align="center" style="background:#f0f0f0;"|'''Total media'''
| align="center" style="background:#f0f0f0;"|'''ul sender'''
| align="center" style="background:#f0f0f0;"|'''ul LB media'''
| align="center" style="background:#f0f0f0;"|'''ul spent ctrl media'''
|-
| 10||1800||180||1620||0
|-
| 5||1800||90||1620||90
|-
| 2.5||1800||45||1620||135
|-
| 1.25||1800||22.5||1620||157.5
|-
| 0.625||1800||11.25||1620||168.75
|-
| 0.3125||1800||5.625||1620||174.375
|}


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Revision as of 14:01, 26 February 2014

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02/26/2014

Spun overnight cultures of GFP-Senders at 13.1g for 10 minutes in 2mL tubes in benchtop centrifuge. Filtered using Fuqing's 0.45μm sterile filters and syringes. Made 5mL of 1% LuxI media. 5mL of LB+amp and 50ul LuxI sender.
Set up microfluidics with David Menn. LuxI inducing LuxR-GFP at 1%.
Made dilutions of filtered sender media. Wanted to make sure each concentration has the same ratio of spent media to fresh media. Made 1800ul. 300ul for 3 replicates of each dilution. 1500ul for flow cytometry. Can take some off the top at multiple time points.

%media Total media ul sender ul LB media ul spent ctrl media
10 1800 180 1620 0
5 1800 90 1620 90
2.5 1800 45 1620 135
1.25 1800 22.5 1620 157.5
0.625 1800 11.25 1620 168.75
0.3125 1800 5.625 1620 174.375


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