Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/05/23: Difference between revisions
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'''Colony PCR on Sender Constructs'''<br> | |||
F primer P093<br>R primer P101<br>RhlI - 10 colonies<br>EsaI - 10 colonies<br>LasI - 10 colonies<br>+ ctrl 1 rxn<br>Positive control is pTet-mCherry 1:1000 miniprepped vector, which should be what a religated backbone is.<br>10μL reactions, annealing T 41°C, extension time 1:30 | |||
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Picked each of the colonies on the receiver plates I made trying to test if any of the constructs were correct. I picked a big swab of each colony and swirled it in some PBS. Then I ran each of them on the flow cytometer. I used the senders (which definitely have GFP) as positive controls. The RhlI sender was green and every. other. colony. was not. This is clearly not a clever and efficient way to screen for receiver constructs. | Picked each of the colonies on the receiver plates I made trying to test if any of the constructs were correct. I picked a big swab of each colony and swirled it in some PBS. Then I ran each of them on the flow cytometer. I used the senders (which definitely have GFP) as positive controls. The RhlI sender was green and every. other. colony. was not. This is clearly not a clever and efficient way to screen for receiver constructs. | ||
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Revision as of 15:52, 29 May 2014
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mm/dd/yyyyColony PCR on Sender Constructs
Picked each of the colonies on the receiver plates I made trying to test if any of the constructs were correct. I picked a big swab of each colony and swirled it in some PBS. Then I ran each of them on the flow cytometer. I used the senders (which definitely have GFP) as positive controls. The RhlI sender was green and every. other. colony. was not. This is clearly not a clever and efficient way to screen for receiver constructs.
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