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

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(Autocreate 2014/02/26 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
Current revision (17:01, 26 February 2014) (view source)
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==02/26/2014==
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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.
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<br>
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Set up microfluidics with David Menn. LuxI inducing LuxR-GFP at 1%.
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<br>
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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.
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''%media'''
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| align="center" style="background:#f0f0f0;"|'''Total media'''
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| align="center" style="background:#f0f0f0;"|'''ul sender'''
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| align="center" style="background:#f0f0f0;"|'''ul LB media'''
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| align="center" style="background:#f0f0f0;"|'''ul spent ctrl media'''
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|-
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| 10||1800||180||1620||0
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|-
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| 5||1800||90||1620||90
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|-
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| 2.5||1800||45||1620||135
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|-
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| 1.25||1800||22.5||1620||157.5
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|-
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| 0.625||1800||11.25||1620||168.75
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|-
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| 0.3125||1800||5.625||1620||174.375
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|}
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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
10180018016200
5180090162090
2.51800451620135
1.25180022.51620157.5
0.625180011.251620168.75
0.312518005.6251620174.375


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