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

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==mm/dd/yyyy==
==mm/dd/yyyy==
{|table
{| {{table}}
|-
|Plasmid||OD650 1||OD650 2||GFP 1||GFP 2
|-
|RhlI||
|-
|EsaI||
|-
|LuxR||
|-
|}
<br>
Added 20ul of each sender and 40ul of LuxR to 20mL LB+amp. Started shaking at 11:15am<br><br>
'''Ligation'''<br>
{| {{table}}
|-
|Bb||6ul||
|-
|Insert||1-5ul
|-
|}
Ligase: 1ul<br>2x buffer 8.5ul<br>Ligation ran for 3 hours at RT<br><br>
Stopped shaking cells at 7PM<br>
:spun senders in 50mL conicals at 200RCF for 10min at 23°C in Wang lab centrifuge<br>
Transferred supernatant to new 50mL conicals<br>Put receivers in 50mL conicals<br>Put all at 4°C.




Revision as of 16:07, 15 November 2013

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mm/dd/yyyy

Plasmid OD650 1 OD650 2 GFP 1 GFP 2
RhlI
EsaI
LuxR


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

Ligation

Bb 6ul
Insert 1-5ul

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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