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

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(mm/dd/yyyy)
Current revision (18:07, 15 November 2013) (view source)
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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.

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mm/dd/yyyy

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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