Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/05/30: Difference between revisions
Rene M Davis (talk | contribs) |
Rene M Davis (talk | contribs) |
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Supernatant transfer: | Supernatant transfer: | ||
Spun down 4ml of culture in 2-2ml tubes for each sender. Poured supernatant into round bottom tubes. Dropped some supernatant onto pH strip. Added 1ml of receiver cells to a 2ml centrifuge tube. Added 20ul of Amp and shook. Added 100ul of Receivers + (~4ul) Amp to each 4ml volume of Sender cells in round bottom tubes. Added 1ml of LB+Amp to add nutrients and lower the pH a little. Same Sender-Receiver pairs as above. Started shaking ~10:30. | |||
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Determine pH using test strips: | Determine pH using test strips: | ||
{| {{table}} | {| {{table}} | ||
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|Sender||pH | |Sender||pH | ||
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| | |Plain LB||6-7 | ||
|- | |- | ||
| | |LuxI||7-8 | ||
|- | |- | ||
| | |LasI||7-8 | ||
|- | |- | ||
| | |RhlI||7-8 | ||
|- | |- | ||
|TraR ctrl|| | |TraI||7-8 | ||
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|TraR ctrl||7-8 | |||
|- | |- | ||
|} | |} | ||
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Revision as of 10:31, 30 May 2013
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Entry title9AM BL21 sender cultures grew in 6mls and 2mls. RhlR-GFP still didn't grow.
Supernatant transfer:
Spun down 4ml of culture in 2-2ml tubes for each sender. Poured supernatant into round bottom tubes. Dropped some supernatant onto pH strip. Added 1ml of receiver cells to a 2ml centrifuge tube. Added 20ul of Amp and shook. Added 100ul of Receivers + (~4ul) Amp to each 4ml volume of Sender cells in round bottom tubes. Added 1ml of LB+Amp to add nutrients and lower the pH a little. Same Sender-Receiver pairs as above. Started shaking ~10:30.
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