IGEM:MIT/2007/Notebook/2007-7-16: Difference between revisions
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#Create Liquid Cultures in LB Media | #Create Liquid Cultures in LB Media | ||
#Dilute Back 1:500 in LB Media | #Dilute Back 1:500 in LB Media | ||
#Grow up for an hour in warm room | #Grow up for an hour in warm room roller | ||
#<font "color=red">Split culture in two</font> | |||
#<font "color=red">Add AHL to one culture to a final concentration of 1E-6M</font> | |||
#<font "color=red">Grow up for 4 hours in warm room roller</font> | |||
#Spin Down, Decant, and Resuspend in PBS | #Spin Down, Decant, and Resuspend in PBS | ||
# | #<font "color=red">Measure absorbance at 600nm and red fluorescence level</font> | ||
=Today's Agenda= | =Today's Agenda= | ||
Revision as of 07:16, 16 July 2007
F2620 Expression Protocol
- Create Liquid Cultures in LB Media
- Dilute Back 1:500 in LB Media
- Grow up for an hour in warm room roller
- Split culture in two
- Add AHL to one culture to a final concentration of 1E-6M
- Grow up for 4 hours in warm room roller
- Spin Down, Decant, and Resuspend in PBS
- Measure absorbance at 600nm and red fluorescence level
Today's Agenda
- Redo Overgrown Liquid Cultures
- Send for Sequencing
- Look for AHL Induced Expression
- Cut CPX and Backbone for ligation
- Start designing high copy backbone
- PCR off relevant portion of Mer operon from pto40
- Organize fridge
- Design and order EC_20 Insert
- Email Isadora + Tom Knight for Meeting Times
Status of Expected Parts
- IDT Primers + Inserts - Arriving Today
- CPX - At FedEx Facility in Boston
- FhuA - ?
