Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/31

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PlasmidOD650 1OD650 2GFP 1GFP 2
RhlI
EsaI
LuxR


Added 20ul of each sender and 40ul of LuxR to 20mL LB+amp. Started shaking at 11:15am

Ligation

Bb6ul
Insert1-5ul

Ligase: 1ul
2x buffer 8.5ul
Ligation ran for 3 hours at RT

Stopped shaking cells at 7PM

spun senders in 50mL conicals at 200RCF for 10min at 23°C in Wang lab centrifuge

Transferred supernatant to new 50mL conicals
Put receivers in 50mL conicals
Put all at 4°C.



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